Process for determining excitants, attractants, stimulants and incitants for members of the class crustacea; method for exciting and/or attracting and/or stimulating and/or inciting members of the class crustacea and feeding compositions

ABSTRACT

Described is a process for determining excitants, attractants, incitants and stimulants for members of the Class Crustacea including but not limited to Penaeus setiferus, Penaeus aztecus, Penaeus vannamei and Procambrus clarkii. The apparatus for carrying out the process includes static tank means, flow-through tank means and Y-maze means each equipped with variable-focus camera means. Also described is a method for exciting and/or attracting and/or stimulating and/or inciting members of the Class Crustacea including but not limited to Penaeus setiferus, Penaeus aztecus and Penaeus vannamei (referred to collectively as Penaeus spp.), as well as Procambrus clarkii, using various dilute aqueous solutions of molecules including but not limited to N-acetyl-D-Glucosamine which is an epimeric mixture of isomers having the structures: ##STR1## as well as feeding compositions for feeding members of the Class Crustacea including (i) prior art feeding compositions and admixed therewith (ii) Crustacea Class-exciting and/or attracting and/or stimulating and/or inciting amounts and concentrations of excitants, attractants, stimulants and/or incitants which include molecules including but not limited to the compounds having the structures: ##STR2##

RELATED CO-PENDING PATENT APPLICATIONS

This is a Divisional of application Ser. No. 08/623,939 filed on Mar.28, 1996 which, in turn, is a Continuation-in-Part of Application forU.S. patent Ser. No. 08/413,440 filed on Mar. 30, 1995, now U.S. Pat.No. 5,579,723 issued on Dec. 3, 1996 which, in turn, is aContinuation-in-Part of Application for U.S. patent Ser. No. 08/279,181filed on Jul. 22, 1994, now U.S. Pat. No. 5,474,030 issued on Dec. 12,1995.

An additional related copending application is Application for U.S.patent Ser. No. 08/535,737 filed on Sep. 28, 1995 (Attorney's Docket No.IFF-9850K) entitled: "PROCESS FOR DETERMINING EXCITANTS, ATTRACTANTS ANDSTIMULANTS FOR MEMBERS OF THE PROCAMBRUS CLARKII SPECIES OF THE CLASSCRUSTACEA, AND METHOD FOR EXCITING AND/OR ATTRACTING AND/OR STIMULATINGMEMBERS OF THE PROCAMBRUS CLARKII SPECIES OF THE CLASS CRUSTACEA". Thisapplication is also a Continuation-in-Part of Application for U.S.patent Ser. No. 08/279,181 filed on Jul. 22, 1994.

BACKGROUND OF THE INVENTION

Our invention is drawn to a process for determining excitants,stimulants, attractants and incitants for members of the Class Crustaceaand a method for exciting, inciting, stimulating and/or attractingmembers of the Class Crustacea by means of placing within a body ofsaline water near a surface or throughout the volume to which suchCrustacea are desired to be attracted, stimulated, incited and/orexcited, a Crustacea attracting, stimulating, inciting and/or excitingconcentration and quantity of at least one of the substances:

(i) N-acetyl-D-Glucosamine, an epimeric mixture of compounds having thestructures: ##STR3## (ii) S-methyl methionine sulfonium halides definedaccording to the structure: ##STR4## (wherein X is chloro or bromo);(iii) methionine having the structure: ##STR5## (iv) trimethyl amineoxide hydrate having the structure: ##STR6## (v) 1-octen-3-o1 having thestructure: ##STR7## (a mixture of isomers having the structures:##STR8## (vi) methional having the structure: ##STR9## (vii) dimethylsulfoxide having the structure: ##STR10## (viii) 50:50 mole:mole mixtureof skatole/indole, skatole having the structure: ##STR11## indole havingthe structure: ##STR12## (ix) propionthetin halides having thestructure: ##STR13## wherein X is chloro or bromo; (x) ammoniumchloride;

(xi) ammonium acetate;

(xii) acetic acid having the structure: ##STR14## (xiii) glucose; (xiv)"raw sugar" (sucrose and "impurities");

(xv) Thaumatin (known as TALIN® a trademark of the Tate and Lyle CompanyLimited of the United Kingdom), a mixture Thaumatin B, Thaumatin I andThaumatin II, the liquid/the liquid chromatograms of which are indicatedin FIGS. 23 and 24 attached hereto and described, infra. Thaumatin I isshown also by the symbol:

     Lys.sup.46, Asp.sup.113 Asp.sup.137 !

as further specifically described in U.S. Pat. No. 5,221,624 issued onJun. 22, 1993 the specification of which is incorporated herein byreference;

(xvi) 2-methyl-3-(methyldithio)furan having the structure: ##STR15##(xvii) aqueous ammonia; (xviii) yeast extract;

(xix) D-Glucosamine, an epimeric mixture of compounds having thestructures: ##STR16## (xx) 2-methyl-3-furanthiol (also known as"OXYCYCLOTHIONE-030™", a trademark of International Flavors & FragrancesInc.) having the structure: ##STR17## (also hereinafter referred to as"ASIE" substance(s)) as well as feeding compositions containing (i) oneor more of such "ASIE" substances admixed with (ii) prior art feedingcompositions.

Aquatic animals utilize water-borne "chemical signals" (chemicalstimuli) to identify and orient toward potential food sources, to escapepredators and locate mates. These specific chemical signals arerecognized in spite of the chemical complexity of aquatic environments.Therefore, the chemical environment of aquatic animals is vitallyimportant, both physiologically and behaviorally, to understand thestatus and role of animals in the aquatic environment. The function ofspecific chemical signals becomes even more significant in a managedbiological system (i.e., aquaculture ponds or tanks) that is optimizedfor production of a single aquatic species (e.g., members of the Penaeusgenus of the Class Crustacea) since these chemical signals regulatefeeding behavior and possibly control reproduction. Because feeds are asignificant expense in most aquaculture operations, the need to maximizefeeding rates and reduce wasted feed, thereby lowering production costsand the possible lowering of bacterial/viral infections is paramount toeconomic success.

The importance of chemoattractants and/or feeding stimulants inimproving both initial palatability and overall feeding rates as a meansto reduce wasted feed is now fully recognized. The feed quality andenvironmental conditions (i.e., water quality and current patterns) havedirect effects on the effectiveness of feed attractants and feedstimulants. For these reasons, food detection and feeding stimulationultimately determine the commercial value of an aquatic feed.

A number of attempts at obtention of efficacious feeding stimulants forvarious aquatic species and for creation of appropriate testingapparatus having a high degree of efficiency for determining goodstimulants and attractants for aquatic species are set forth in theliterature. Thus, U.S. Pat. No. 4,250,835 issued Feb. 17, 1981 and U.S.Pat. No. 4,249,480 issued Feb. 10, 1981 disclose apparatus and methodsfor rearing shrimp through the larvae stage wherein the shrimp aresubjected to controlled conditions and a common enclosure for the maleand female adult shrimp is provided which permits uncontrolled access ofthe shrimp to one another and wherein the shrimp are maintained througha plurality of cycles of mating, spawning and hatching. The systemdisclosed provides filtration means for filtering the medium of thecommon enclosure and with collecting means for harvesting hatched shrimpat preselected times from the common enclosure medium as the mediummoves into the filtration means. U.S. Pat. No. 4,828,829 of May 9, 1989discloses a visual fish attractant that aids in the dispersion oftraditional scent and taste attractants. The fish attractantcompositions include one or more oils, such as mineral oil, cod liveroil, menhaden oil, herring oil, anise oil, salmon oil, as well aspigments, fragrances, fish scent, dispersed pigments, andlight-reflective particles that act both as a visual attractant and asan aid to controlled dispersion of the oil and scent composition.

Lombardo, et al, Comp.Biochem.Physiol., Vol. 101C, No. 2, pages 389-398,1992, "Amino Acids and Derivatives as Food-Finding Signals in theFreshwater Snail Planorbarius corneus (L.)" discloses the behavioralresponses of the freshwater snail to various amino acids including1-aspartic acid, d-alanine, histamine, proline and aspartame.

It should be pointed out, however, that the compounds having thestructures: ##STR18## wherein X is a "univalent anion" such as chlorideion or bromide ion are used to attract in a gel vertebrate fish such asred snapper and carp in Japanese Published Application J91/27231(Nakajima) abstracted at Chem. Abstracts, Volume 115:113303n. Thedisclosure of Japanese Published Application 91/27231 does not detractfrom the patentability of the instant invention.

Nothing in the prior art, however, discloses the efficient process forattracting, inciting, stimulating and/or exciting members of the ClassCrustacea from or in a volume of water inhabited by said member(s) ofthe Class Crustacea in or to a desired location or volume within a bodyof water by applying at least one of the specific materials found to beuseful in our invention, to wit the "ASIE" substances found to be souseful in our invention. Furthermore, nothing in the prior art disclosesfeeding compositions containing such "ASIE" substances admixed withprior art feeding materials for such members of the Class Crustacea.

THE INVENTION

Our invention provides a process for attracting, exciting, stimulatingand/or inciting at least one member of the Class Crustacea from a volumeof water or a surface inhabited by said member of the Class Crustacea toa desired location or volume within a body of water comprising the stepof applying an aqueous saline solution containing aCrustacea-attracting, exciting, stimulating and/or incitingconcentration of at least one substance (referred to as "ASIE"substance(s)) selected from the group consisting of:

(i) N-acetyl-D-Glucosamine, an epimeric mixture of compounds having thestructures: ##STR19## (ii) at least one S-methyl methionine sulfoniumhalide having the structure: ##STR20## (wherein X is a chloride anion ora bromide anion); (iii) methionine having the structure: ##STR21## (iv)trimethyl amine oxide hydrate having the structure: ##STR22## (v)(R)(S)1-octen-3-o1 having the structure: ##STR23## (a mixture of isomershaving the structures: ##STR24## (vi) methional having the structure:##STR25## (vii) dimethyl sulfoxide having the structure: ##STR26##(viii) 50:50 mole:mole mixture of skatole/indole, skatole having thestructure: ##STR27## indole having the structure: ##STR28## (ix)propionthetin halides having the structure: ##STR29## wherein X is achloride anion or a bromide anion; (x) ammonium chloride;

(xi) ammonium acetate;

(xii) acetic acid having the structure: ##STR30## (xiii) glucose; (xiv)raw sugar (sucrose and "impurities") for example, Osceola Brown Sugarmarketed by Osceola Farms Inc. of Pahokee, Fla. (the headspace analysisfor which is set forth in FIG. 27 described in detail, infra);

(xv) Thaumatin (known as TALIN® a trademark of the Tate and Lyle CompanyLimited of the United Kingdom), a mixture Thaumatin B, Thaumatin I andThaumatin II, the liquid/the liquid chromatograms of which are indicatedin FIGS. 23 and 24 attached hereto and described, infra. Thaumatin I isshown also by the symbol:

     Lys.sup.46, Asp.sup.113, Asp.sup.137 !

as further specifically described in U.S. Pat. No. 5,221,624 issued onJun. 22, 1993 the specification of which is incorporated herein byreference;

(xvi) 2-methyl-3-(methyldithio)furan (also called "MOS" or"METHYLOXYCYCLOSULFIDE™", a trademark of International Flavors &Fragrances Inc.) having the structure: ##STR31## (xvii) aqueous ammonia;(xviii) yeast extract;

(xix) D-Glucosamine, an epimeric mixture of compounds having thestructures: ##STR32## (xx) 2-methyl-3-furanthiol (also known as"OXYCYCLOTHIONE-030™", a trademark of International Flavors & FragrancesInc.) having the structure: ##STR33## taken alone, or in combinationwith naturally occurring excitants, stimulants, attractants and/orincitants for the Class Crustacea such as natural Crustacean/cephalopodextracts to the vicinity of said desired location or volume. Ourinvention also describes a process for exciting, attracting, stimulatingand/or inciting a member of the Class Crustacea within a volume of waterinhabited by such member of the Class Crustacea comprising the step ofapplying an aqueous solution containing a Crustacean-exciting,attracting, inciting and/or stimulating concentration of one of theabove materials (designated as "ASIE" materials) to the vicinity of saidvolume inhabited by said member of the Class Crustacea.

Examples of such members of the Class Crustacea are members of thePenaeus genus of the Class Crustacea such as Penaeus setiferus, Penaeusvannamei and Penaeus aztecus; and the Procambrus clarkii species of theClass Crustacea

The ultimate goal of the ascertainment of such attractancy, stimulation,incitance and/or excitement is to intensify the feeding kinetics (e.g.,rate and quantity of food intake and metabolism) for the members of theClass Crustacea. Accordingly, this invention also pertains to a feedingprotocol for such members of the Class Crustacea relating to theaforesaid ascertainment and feeding compositions pertaining to saidfeeding protocol.

Thus, our invention is also directed to feeding compositions for feedingmembers of the Class Crustacea including (i) prior art feedingcompositions (such as RANGEN®-35, a mixture of 35% protein, 7% fat, fishmeal, fish oil and minor amounts of mineral and vitamin nutrients) andalso containing, for example, L-ascorbic acid-2-phosphate or its saltsas described in Published Japanese Applications J94/093,821 andJ94/093,822 assigned to Showa Denko KK and abstracted in the DerwentPatents Index Alerting Abstracts Bulletin for week 9445 issued on Jan.4, 1995 as follows:

SHOW D13 87-253800/36=JP94093821.B2 Food additive for fishfarming.contains L-ascorbic acid-2-phosphate or its salts as activeingredient

SHOWA DENKO KK 86.01.30 86JP.016739

C01 (94.11.24) *JP62175142-A A23K 1/18, 1/16, A61K 31/375//C07D 307/62

Based on JP62175142A

A food additive contg. L-ascorbic acid-2-phosphate or its salts asactive ingredient is claimed.

USE/ADVANTAGE--Used as a stable food additive with an ascorbic acideffect for fish farming.

In an example, a pellet fodder (basic fodder) composed of 55% white baltpowder, 32% alpha-potato starch, 5% soy bean oil, 3% McCollum and 5%ascorbic acid contg. vitamins was prepd. This was used in an ascorbicacid shortage fodder zone. To the basic fodder compsn. (1 kg), 2 m.molof ascorbic acid was added for use in an ascorbic acid added fodderzone. To the basic fodder compsn. (1 kg), 2 m.mol of L-ascorbicacid-2-phosphate Mg was also added. The vitamins mixt. (5 g) contained 6mg of thiamine, HCl. 20 mg of riboflavin, 4 mg of pyridoxic HCl, 0.009mg of vitamin B12, 80 mg of nicotinic acid, 800 mg of Chlorin chloride,400 mg of inositol, 28 mg of ca-pantothenate, 6 mg of biotin, 1.5 mg ofacid 1.5 mg. 40 mg alphatocopherol, 4 mg menadione, 0.05 mg ofcalciferol, 20 mg of letinene.acetate and 3596 mg of cellulose. Satd.pellet fodders were suspended to the fry of rainbow trout which were fedfor 100 days. In each zone, 200 of fries were fed. The fish weresupplied 6 times per day from the first until the 80th day and 4 timesper day from the 81st until the 100th day. The difference of averagebody wt. between zones was little, but transformed fries and mortalitywere decreased in the L-ascorbic acid adding zone. No appearance oftransformed fries were observed in the L-ascorbic acid-2-phosphate Mgadded zone. (5 pp)

SHOW D13 88-025158/04=JP94093822.B2 Ingredient for feed ofcrustacea-comprises ester of L-ascorbic acid-2-phosphoric acid or itssalt

SHOWA DENKO KK 86.06.05 86JP-129283

C03 (94.11.24) *JP62285759-A A23K 1/18, 1/16, A61K 31/375

Based on JP62285759A

The ingredient comprises an appropriate amt. of the ester of L-ascorbicacid-2-phosphoric acid or its salt, in the feed.

USE/ADVANTAGE--A nutritious and economical ingredient used as anefficient growth promoter for e.g. shrimps or lobsters. (6 pp) and/orfish growth hormone polypeptides produced by culturing microorganismscarrying recombinant DNA plasmids as described in Published JapaneseApplication J94/095,938 assigned to Kyowa Hakko Kogyo KK and abstractedas set forth below:

KYOW D16 86-008665/02=JP94095938-B2 Fish growth hormonepoly-peptide-produced by culturing microorganisms carrying recombinantDNA plasmid

KYOWA HAKKO KOGYO KK 85.03.13 92JP-204982 (Div ex

85.03.13 85JP-050096)

C03 (94.11.30) *EP 166444-A C12N 15/18, C07K 13/00,

C12N 1/21//A23K 1/165, A61K 35/74, 37/36, C12P 21/02 (C12N 1/21, C12R

1:19) (C12P 21/02, C12R 1:19)

Based on JP5207883-A

Addnl. Data; SEKINE S (SEKI/)

Fish growth hormone polypeptide (I) having a disclosed peptide sequenceis new. Also claimed are DNA and recombinant DNA which code for (I) andmicroorganisms contg. the recombinant DNA which codes for (I).

An m-RNA of fish growth hormone is isolated from the pituitary gland offish belonging to Cupleiformes e.g. salmon, and used as a template toprepare cDNA. A recombinant plasmid (e.g. pSGHI and PSGH14)incorporating the cDNA is prepd. The recombinant plasmid is incorporatedin a host microorganism. The DNA and recombinant plasmid can be used forexpression of fish growth hormone gene in bacteria such as E.coli. Fishgrowth hormones are produced by culturing microorganisms carrying therecombinant plasmid.

USE--(I) has a stimulating effect on the growth of fish and is used as acomponent of bait for fish cultivation. (11 pp)

as published in the Derwent Chemical Patents Index Alerting AbstractsBulletin for week 9501 issued on Feb. 3, 1995 and being admixedtherewith (ii) Crustacea-Class exciting and/or attracting and/orinciting and/or stimulating amounts and concentrations of at least oneof the substances set forth as "ASIE" substances, supra.

Useful in practicing our invention is apparatus for determining whethera given substance at a given aqueous concentration or variableconcentrations attracts, excites, incites and/or stimulates at least onemember of the Class Crustacea comprising static holding tank means,flow-through holding tank means and/or Y-maze apparatus means.

More specifically, our invention is practiced using apparatus fordetermining whether a given substance at a given aqueous concentrationor variable concentrations excites a member of the Class Crustaceacomprising:

(a) static holding tank means containing (i) a volume of water and (ii)at least one live member of the Class Crustacea within said volume ofsaline water;

(b) pump-injected test solution feeding means for pumping aqueoussolutions at variable or constant flow rates of test substance into thestatic holding tank means;

(c) visible light generating and guidance means for guiding visiblewavelength light of variable or preferably constant intensity into saidvolume of water in said static holding tank means;

(d) first variable power source means for engaging the visible lightgenerating means;

(e) second variable power source means for engaging the pump generatedfeeding means; and

(f) variable focus/focal length video camera or camcorder recordingmeans for recording the movements of one or more specified bodyappendages and/or the lateral direction and velocity of one or morespecified body appendages of one or more of the members of the ClassCrustacea when the pump injected feeding means and visible lightgenerating means are engaged;

whereby it can be determined whether a particular dilution of a testsubstance will cause a member of the Class Crustacea to be "excited".

The "recording" by the variable focus/focal length video camera orcamcorder recording means is preferably for recording on film ormagnetic tape.

An additional aspect of the apparatus useful in practicing our inventionis covered herein for determining whether a given substance at a givenaqueous concentration or variable concentrations will attract, excite orincite members of the Class Crustacea. Such apparatus comprises:

(a) flow-through holding tank means where varying concentrations of testsolutions are fed in at the inlet of the holding tank means and flowthrough the holding tank to an exit portal while a member of the ClassCrustacea is positioned within the "flow-through" holding tank. Theholding tank contains (i) flowing saline water and (ii) at least onelive member of the Class Crustacea within the flowing saline water;

(b) pump-injected test solution feeding means for pumping aqueoussolutions at variable or preferably constant flow rates of test solutioninto an orifice in front of the entry portal into the flow-throughholding tank;

(c) visible light generating and guidance means for guiding visiblewavelength light of variable or preferably constant intensity into thevolume of water flowing in the flow-through holding tank;

(d) a first variable power source for energizing or "engaging" thevisible light generating means; (e) a valuable power source forenergizing or "engaging" the pump injection feeding means (the pump hasvariable heads so that the flow rate of the added chemical solution(containing at least one "ASIE") may be added to the basic flow ofsaline water through the flow-through holding tank);

(f) variable focal length video camera or camcorder recording means forrecording the movements and/or the lateral direction and velocity of oneor more members of the Class Crustacea when the pump injection feedingmeans is engaged and when the visible light generating means is engaged;and

(g) specially designed camera platform and blind means (a part of ourinvention) the purpose of which is to shield the observer from the testanimal in order to prevent the test animal from being extraneouslystressed and/or distracted. (The variable focal length video camera orcamcorder recording means is mounted within the specially designedcamera platform and blind means.)

The variable focal length video camera or camcorder recording means isof necessity on a line of visibility between the camera lens and thelimits of motion of the member of the Class Crustacea. Accordingly, theapparatus useful in practicing our invention must contain one or twoclear panels behind which is located the video camera or camcorder andin front of which is located at every point of view of the lens of thevideo camera or camcorder all of the members of the Class Crustaceawithin the flow-through holding tank or within the static holding tankor within the Y-maze apparatus as the case may be.

More specifically, the variable focus multi-directional video camera orcamcorder/video camera or camcorder platform apparatus for facilitatingvisible wavelength radiation communication between the moving member ofthe Class Crustacea and an image display of the moving object on, forexample, visible radiation sensitive film or visible radiation sensitivemagnetic tape comprising:

(1) a horizontal static substantially rectangular planar base located inan "X-Y" plane, having an upper surface and having two substantiallyparallel ends parallel to the "X" axis and two substantially parallelends parallel to the "Y" axis; with one end parallel to the "Y" axisbeing along a line designated "E₁ ";

(2) a vertically disposed substantially rectangular planar frame locatedin a "Y-Z" plane, substantially perpendicular to said horizontal staticplanar base and having two parallel ends parallel to the "Y" axis andtwo parallel ends parallel to the "Z" axis, one of said ends parallel tothe "Y" axis being contiguous with said static planar base along a firstend, said line E₁ of said static planar base, parallel to the "Y" axis,said frame having a first orifice therethrough permitting visiblewavelength radiation to be transmitted therethrough;

(3) a continuous right angle-shaped horizontal/vertical planarbase/frame movable platform movable in both the "X" and "Y" (or lateral)directions consisting of (i) a substantially rectangular horizontallamina located in the "X-Y" plane facing and within the planar frameworkof said horizontal static planar base (1) plane and having an uppersurface and a lower surface continuously and sealably juxtaposed in adirection parallel to line E₁ along a line designated E₂ said line E₂being proximate said line E₁ with (ii) a substantially rectangularvertical lamina located in the "Y-Z" plane facing and within the planarframework of said vertically disposed frame (2) and containing avertically disposed visible wavelength radiation shield (blind) having asecond orifice therethrough located opposite to and being planarlyparallel to and within the planar framework of said first orifice insaid vertically disposed frame, said right angle-shaped movable platformbeing located on rollers juxtaposed with and immediately adjacent saidlower surface of said right angle-shaped movable platform and said uppersurface of said horizontal static base;

(4) a substantially rectangular rotating planar base having an uppersurface and a lower surface and having two parallel ends parallel to the"Y" axis and two parallel ends parallel to the "X-Z" plane, beingrotatable about a vertex line E₃ in a hinge manner having a directionparallel to each of lines E₁ and E₂ and being substantially proximatelines E₁ and E₂ along a first parallel hinge end in the "Y" directionparallel to the "Y" axis, having variable manually-controllable rotationmovement means located at an end of said rotating planar base oppositesaid first parallel hinge end, and opposite the location of said lineE₃, the direction of rotation of said rotating base being from thelocation of the "X-Y" plane of said right angle-shaped movable platformtowards the location of the "Y-Z" plane of said right angle-shapedmovable platform about the said vertex line E₃ ; and

(5) a video camera or camcorder fixedly mounted on the upper surface ofsaid rotating planar base, comprising a housing and a lens located insaid housing, the focal plane of said lens being substantially parallelto and at a controllably variable distance from the plane of said firstorifice of said vertically disposed frame and said second orifice ofsaid visible wavelength radiation shield with said lens, said firstorifice and said second orifice all having a common line of vision,

whereby simultaneous manipulation of said rotating planar base and saidright angle-shaped movable platform, when the video camera or camcorderis in operation, facilitate visible wavelength radiation communicationbetween (i) an image display resulting from operation of said videocamera or camcorder onto, for example, visible light sensitive film orvisible light sensitive magnetic tape and (ii) the image of the movingobject (that is the member of the Class Crustacea) being transmitted tosaid video camera or camcorder.

Another aspect of apparatus useful in the practice of our inventionconcerns an additional apparatus for determining whether a givensubstance at a given aqueous concentration or variable concentrationsattracts, or stimulates to feed, or incites to feed, or excites, orstimulates a member of the Class Crustacea; and it comprises a "Y-maze"apparatus similar to that disclosed in a paper by Lee,J.Exp.Mar.Biol.Ecol., Volume 153 (1992), pages 53-67, "Chemotaxis byOctopus maya Voss et Solis in a Y-maze".

More specifically, an aspect of apparatus useful in the practice of theinstant invention comprises, inter alia, a recirculating tank meanscontaining a Y-maze which is a rectangular parallelepiped as its"three-dimensional base" having two rectangular parallelepiped-shapedarms; the three-dimensional base having one closed end and one open endand two sides, each sealably connected to an end panel and eachterminating at the open end, said open end connected to two divergingparallelepiped-shaped sections as set forth, supra (having a vertex of5°-45° (preferably having a 100 vertex)); (i) a test substance inputsection and (ii) a control section, each of which is sealably connectedat the open end to one another and to a side of the firstthree-dimensional base rectangular parallelepiped section. Thisapparatus has one or both top and bottom panels as transparent panelswhereby a variable focal length video camera or camcorder recordingmeans is maintained on one side of the transparent panel with a line ofvision from the lens of the video camera or camcorder to every placewhere all of the members of the Class Crustacea may travel. It should benoted that the control section and test substance input section can bereversed or interchanged (as by changing the location of the testsubstance input tube as more particularly shown in FIG. 5G and thedetailed description of FIG. 5G in the "Detailed Description of theDrawings" section, infra).

In the Y-maze apparatus useful in the practice of our invention, aconstant concentration of the test chemical entering one arm of theY-maze is the preferred method of addition. In the flow-throughapparatus, however, there is a constant increase of concentration of thetest chemical during the run.

Furthermore, the Y-maze apparatus useful in the practice of ourinvention also contains a volume of water and at least one live memberof the Class Crustacea within the volume of water. The Y-maze apparatususeful in the practice of our invention contains a pump-injected testsolution of test chemical for pumping aqueous solutions of test chemicalat various flow rates or preferably constant flow rates into the feedingsections of the Y-maze (at one of the diverging parallelepiped-shaped"arms" connected to the base parallelepiped).

Like the flow-through apparatus and the static tank apparatus, the"Y-maze" apparatus has a visible light generating and guidance means forguiding visible wavelength light of variable or preferably constantintensity into the volume of water in the holding tank and feedingsection where a member of the Class Crustacea is located.

The "Y-maze" aspect of the apparatus useful in the practice of ourinvention also contains variable focal length video camera or camcorderrecording means for recording the movements of one or more appendagesand/or the lateral direction and velocity of one or more of the membersof the Class Crustacea when the pump injection test chemical means andwhen the visible light generating means are simultaneously energized or"engaged".

More specifically, in determining a value for the response, "R" or "G"of the member of the Class Crustacea whose responses are measured as aresult of the practice of our invention, the time taken for the memberof the Class Crustacea to act or to respond to the feeding incitant orstimulant or attractant of our invention is a function of the particularmaterial used and its concentration as well as the flow rate of theliquid.

Proposed mathematical models are set forth herein, to wit: ##EQU1##wherein the term: θis the time taken and the terms "G₁ " and "G₂ " areeach values for the responses of the members of the Class Crustacea fromthe initial time "0" of substance injection until the time that a givendefinitive Crustacean forward motion commences.

It should be noted that the term "G" is also shown herein as "R".

The rate of change of response with respect to test substanceconcentration (C) (for example, in terms of moles per liter or grams perliter) is given by the equation: ##EQU2## wherein the symbol: ##EQU3##is the rate of change of time with respect to concentration, that is,time of response and the symbol: ##EQU4## is the rate of change ofresponse with respect to concentration as is the symbol: ##EQU5## Asimilar equation is: ##EQU6## which depends on the mathematical model:##EQU7## Combining the initial mathematical model with the differentialequation yields the differential equation: ##EQU8## and the differentialequation: ##EQU9##

Changes in response value when concentrations of stimulant change orwhen concentrations of excitant change or when concentrations ofattractant change are shown by the equations: ##EQU10##

Thus, for example, in the case of the use with the Penaeus vannameispecies of the Pennaeus genus of the Class Crustacea ofN-acetyl-D-Glucosamine, an epimeric mixture of compounds having thestructures: ##STR34## the response versus concentration can be shown bythe equation:

    -log.sub.10 C=0.8 G-2.3

in the static tank, or by one of the equations: ##EQU11## Furthermore,the rate of change of response with respect to concentration can beshown by the equations: ##EQU12## and the change in response can beshown by the equation: ##EQU13## Combining the equation:

    G.sub.1 =2.9-2.9 log.sub.e C.

with the equation: ##EQU14## and the equation: ##EQU15## will yield theequation: ##EQU16## (showing concentration as a function of time; when atest chemical is added continuously as when using the flow-through orstatic apparatus; but not the Y-maze apparatus) and the equation:##EQU17##

When using the flow-through apparatus and using with the Penaeusvannamei species of the Pennaeus genus of the Class Crustacea,N-acetyl-D-Glucosamine, an epimeric mixture of compounds having thestructures: ##STR35## as an excitant, attractant, stimulant and/orincitant (on a scale of 0-50, as opposed to a scale of 1-5 when usingthe static tank-containing apparatus), the mathematical models for themean response as a function of concentration of theN-acetyl-D-Glucosamine, the test chemical, are as follows: ##EQU18## Themathematical model for the median response as a function ofconcentration of N-acetyl-D-Glucosamine, the test chemical, is asfollows:

    R=-41.98-5.68 log.sub.10 C.

Other materials which were tested but which have not been made a part ofour invention are as follows:

(i) Taurine, the compound having the structure: ##STR36## (ii)Guanidine, the compound having the structure: ##STR37## (iii) Betaine,the compound having the structure: ##STR38## (iv) citric acid, thecompound having the structure: ##STR39## (v) Pyruvic acid, the compoundhaving the structure: ##STR40## (vi) Ferulic acid, the compound havingthe structure: ##STR41## (vii) Vanillin, the compound having thestructure: ##STR42## (viii) Isovaleraldehyde, the compound having thestructure: ##STR43## (ix) trans-2-hexenal, the compound having thestructure: ##STR44## (x) Citral, the mixture of compounds having thestructures: ##STR45##

Each of the substances of our invention tested is diluted in a salinesolution containing 30 parts per thousand of a "FRITZ®" Super Saltconcentration manufactured by the Fritz Chemical Company of Dallas, Tex.75217. The FRITZ® Super Salt Concentrate contains in major amountssodium chloride, magnesium sulfate, magnesium chloride and calciumchloride and in minor amounts lithium chloride, sodium molyedate,disodium phosphate, strontium chloride, potassium chloride, sodiumbicarbonate, calcium carbonate and magnesium carbonate.

The volume of water described, supra, for containing a member of theClass Crustacea is a dilute saline solution containing the sameconcentration of 30 parts per thousand of a FRITZ® Super Salt solutioncontaining the above ingredients.

The protocols for using the apparatus of our invention and fordetermination of the feed compositions of our invention are set forth asfollows:

STATIC TANK: TESTING PROTOCOL

I. PREPARATION

1. APPARATUS

A. BETWEEN CHEMICALS

1. Soak test chamber (static tank) in warm water for at least one hourbefore testing different chemicals.

2. Rinse with hot tap water for approximately 30 seconds.

3. Rinse with de-ionized water for 10 seconds.

4. Dry off outside of chamber and place on test stand.

5. Replace peristaltic pump tubing before testing different chemicals.

6. Calibrate peristaltic pump to deliver at the predetermined flow rate.

7. Place tubing inflow and outflow ends into test chemical bottle.

8. Turn on peristaltic pump to fill tube with water and purge out allthe air.

9. Adjust lighting for even illumination in the tank.

10. Set up camera in an appropriate area for viewing the entire bottomof tank.

11. Pour 1,000 mLs of sea water taken from the shrimp holding systeminto the test tank.

B. BETWEEN TRIALS USING THE SAME CHEMICALS

1. Rinse test tank with warm tap water for approximately 30 seconds.

2. Rinse with de-ionized water for 10 seconds.

3. Dry off outside of chamber and place on tank stand.

4. Between the trials using the same chemical or concentration of thesame chemical, pump peristaltic pump tubing with at least 250 mLs saltwater.

5. Calibrate peristaltic pump to deliver at the predetermined flow rate.

6. Place tubing inflow and outflow ends into test chemical bottle.

7. Turn on peristaltic pump to fill tube with water and purge out allthe air.

8. Adjust lighting for even illumination in the tank.

9. Set up camera in an appropriate area for viewing the entire bottom ofthe tank.

10. Pour 1,000 mLs of sea water taken from the shrimp holding systeminto the test tank.

2. TEST ORGANISM

Animals should be chosen from a previously isolated set of animals sorepetition does not occur within the test set for a particular chemicalconcentration. All test animals should be free of chitinolytic bacteriaif possible. Most importantly, animals should have all head and mouthappendages (antennules, antennae, maxillae, maxillipeds and walkinglegs).

1. Select an animal that appears to be calm (i.e., not repetitivelyhitting head into wall).

2. Net the animal carefully.

3. Place the animal into the test tank.

4. Quickly set the cover over the test tank so the animal does not jumpout.

5. If an animal jumps out of the tank or net at any time duringtransfer, pick it up and place it back into the holding chamber andchoose another animal.

II. TEST PROCEDURE

1. PRETRIAL

1. When the animal is placed into the test tank, begin timing theacclimation period of 15 minutes.

2. Record the test date onto the data sheet.

3. When the acclimation period ends, record the acclimation time andturn on the camera to record animal movements.

2. TRIAL

4. Immediately begin timing the run.

5. Observe and record on data sheet the placement and movements of theanimal. Also note any problems that occurred during testing.

6. When the 3 minute test is over, turn off the peristaltic pump, cameraand timer.

3. POST-TRIAL

7. Remove the chemical delivery tube from the tank and place into thetest chemical bottle.

8. Pour the water from the test tank through a net into a waste bucket.The animal should now be in the net.

9. Place the animal into a group or individual holding chamber for latertesting.

10. Begin preparation of apparatus and test animal.

FLOW-THROUGH TANK: TESTING PROTOCOL

I. PREPARATION

1. APPARATUS

A. BETWEEN CHEMICALS

1. Soak test chamber (flow-through tank) and divider in an alkalinesolution and warm water for at least one hour before testing differentchemicals.

2. Rinse with hot tap water for approximately 30 seconds.

3. Rinse with de-ionized water for 10 seconds.

4. Dry off outside of chamber and place on table.

5. Insert the tank divider into the appropriate groove in the center ofthe tank.

6. Replace peristaltic injection pump tubing before testing differentchemicals.

7. Calibrate peristaltic injection pump to deliver at the predeterminedflow rate.

8. Place smaller tubing inflow and outflow ends into test chemicalbottle.

9. Attach larger recirculation tubing to the ends of the tank via thereducers. The flow of water should go from the tank end with thechemical inlet to opposite end of the tank.

10. Turn on peristaltic injection pump to fill tubes with water andpurge out all the air.

11. Adjust lighting for even illumination in the tank.

12. Set up camera blind and camera in an appropriate area for viewingthe entire test tank.

13. Pour 500 mLs of sea water taken from the shrimp holding system intothe test tank.

B. BETWEEN TRIALS USING THE SAME CHEMICAL

1. Rinse test tank and tank divider with warm tap water forapproximately 30 seconds.

2. Rinse with de-ionized water for 10 seconds.

3. Dry off outside of chamber and place on table.

4. Insert the tank divider into the appropriate groove in the center ofthe tank.

5. Between the trials using the same chemical or concentration of thesame chemical, pump peristaltic injection pump tubing with at least 1liter salt water.

6. Calibrate peristaltic injection pump to deliver at the predeterminedflow rate.

7. Place smaller tubing inflow and outflow ends into test chemicalbottle.

8. Attach larger recirculation tubing to the ends of the tank via thereducers. The flow of water should go from the tank end with thechemical inlet to opposite end of the tank.

9. Turn on peristaltic injection pump to fill tubes with water and purgeout all the air.

10. Adjust lighting for even illumination in the tank.

11. Set up camera blind and camera in an appropriate area for viewingthe entire test tank.

12. Pour 500 mLs of sea water taken from the shrimp holding system intothe test tank.

2. TEST ORGANISM

Animals should be chosen from a previously isolated set of animals sorepetition does not occur within the test set for a particular chemicalconcentration. All test animals should be free of chitinolytic bacteriaif possible. Most importantly, animals should have all head and mouthappendages (antennules, antennae, maxillae, maxillipeds and walkinglegs).

1. Select an animal that appears to be calm (i.e., not repetitivelyhitting head into wall).

2. Net the animal carefully.

3. Place the animal into the test tank in the end nearest the tankoutflow, facing the inflow (face the animal against the future flow ofwater).

4. Quickly set the cover over the test tank so the animal does not jumpout.

5. If an animal jumps out of the tank or net at any time duringtransfer, pick it up and place it back into the holding chamber andchoose another animal.

II. TEST PROCEDURE

1. PRETRIAL

1. When the animal is placed into the test tank, begin timing theacclimation period. The duration of the acclimation period should be theperiod of time it takes for the animal to become calm (i.e., not bumpinginto walls and not attempting to swim or turn in tank). This periodshould be at least one minute.

2. Record the test date onto the data sheet.

3. Turn on peristaltic injection pump to allow water to circulate in thetest chamber.

4. When the acclimation period ends, record the acclimation time andturn on the camera to record animal movements.

2. TRIAL

5. Immediately begin timing the run.

6. Carefully remove the tank divider.

7. Wait 5 seconds. If the animal lunges to the inlet side of the testtank during this time, terminate testing.

8. If the animal stays on the outlet end of the tank, place the chemicaloutlet tube into the test chemical inlet of the tank.

9. Wait 30 seconds. If the animal lunges to the inlet side of the testtank during this time, terminate testing.

10. Observe and record on data sheet the placement and movements of theanimal. Also note any problems that occurred during testing.

11. When the predetermined test time is over, turn off the peristalticinjection pump, camera and timer.

3. POST-TRIAL

12. Remove the chemical delivery tube from the chemical delivery inletand place into the test chemical bottle.

13. Remove large recirculation tubing and begin cleaning (see ApparatusPreparation above).

14. Pour the water from the test tank through a net into a waste bucket.The animal should now be in the net.

15. Place the animal into a group or individual holding chamber forlater testing.

16. Begin preparation of apparatus and test animal.

Y-MAZE: TESTING PROTOCOL

I. PREPARATION

1. APPARATUS

A. BETWEEN CHEMICALS

1. Soak test chamber (Y-maze) and divider in an alkaline solution andwarm water for at least one hour before testing different chemicals.

2. Rinse with hot tap water for approximately 30 seconds.

3. Rinse with de-ionized water for 10 seconds.

4. Dry off outside of chamber and place on table.

5. Insert the appropriate tank dividers in the Y-maze.

6. Replace peristaltic injection pump tubing before testing differentchemicals.

7. Calibrate peristaltic injection pump to deliver at the predeterminedflow rate.

8. Place tubing inflow and outflow ends into test chemical bottle.

9. Turn on peristaltic injection pump to fill tube with liquid and purgeout all the air.

10. Adjust lighting for even illumination in the tank.

11. Set up camera in an appropriate area for viewing the entire Y-maze.

12. Fill Y-maze to the proper water level with 5 liters sea water takenfrom the shrimp holding system.

B. BETWEEN TRIALS USING THE SAME CHEMICAL

1. Rinse Y-maze and divider with warm tap water for approximately 30seconds.

2. Rinse with de-ionized water for 10 seconds.

3. Dry off outside of chamber and place on table.

4. Insert the appropriate tank dividers in the Y-maze.

5. Between the trials using the same chemical or concentration of thesame chemical, pump peristaltic injection pump tubing with at least 250mLs salt water.

6. Calibrate peristaltic injection pump to deliver at the predeterminedflow rate.

7. Place smaller tubing inflow and outflow ends into test chemicalbottle.

8. Turn on peristaltic injection pump to fill tube with liquid and purgeout all the air.

9. Adjust lighting for even illumination in the Y-maze.

10. Set up camera in an appropriate area for viewing the entire Y-maze.

11. Fill Y-maze to the proper water level with 5 liters of sea watertaken from the shrimp holding system.

12. TEST ORGANISM

Animals should be chosen from a previously isolated set of animals sorepetition does not occur within the test set for a particular chemicalconcentration. All test animals should be free of chitinolytic bacteriaif possible. Most importantly, animals should have all head and mouthappendages (antennules, antennae, maxillae, maxillipeds and walkinglegs).

1. Select an animal that appears to be calm (i.e., not repetitivelyhitting head into wall).

2. Net the animal carefully.

3. Place the animal in the appropriate position in the Y-maze (in thebase of the Y-maze).

4. If an animal jumps out of the tank or net at any time duringtransfer, return it to the holding chamber and choose another animal.

II. TEST PROCEDURE

1. PRETRIAL

1. When the animal is placed into the Y-maze, turn on the recirculationlines of pump; begin timing the acclimation period of 15 minutes.

2. Record the test date onto the data sheet.

3. When the acclimation period ends, turn on the camera to record animalmovements.

2. TRIAL

4. Immediately begin timing the run and begin pumping test chemical intothe chemical inlet of the Y-maze.

5. After 3 minutes, carefully remove any tank divider.

6. Wait 30 seconds. If the animal lunges into an arm of the Y-mazeduring this time, terminate testing.

7. Observe and record on data sheet the placement and movements of theanimal. Also note any problems that occurred during testing.

8. When the predetermined test time is over, turn off the peristalticinjection pump, camera and timer.

3. POST-TRIAL

9. Return chemical delivery tubes to the test chemical bottles.

10. Remove Y-maze recirculation tubing and begin cleaning (see ApparatusPreparation above).

11. Pour the water from the Y-maze through a net into a waste bucket.The animal should now be in the net.

12. Place the animal into a group or individual holding chamber forlater testing.

13. Begin preparation of apparatus and test animal.

FEEDING BEHAVIOR: TESTING PROTOCOL

I. PREPARATION

1. APPARATUS

A. BETWEEN CHEMICALS

1. Soak test chamber (Y-maze) and divider in an alkaline solution andwarm water for at least one hour before testing different chemicals.

2. Rinse with hot tap water for approximately 30 seconds.

3. Rinse with de-ionized water for 10 seconds.

4. Dry off outside of chamber and place on table.

5. Insert the appropriate tank dividers into the Y-maze.

6. Replace peristaltic injection pump tubing before testing differentchemicals.

7. Calibrate peristaltic injection pump to deliver at the predeterminedflow rate.

8. Place tubing inflow and outflow ends into test chemical bottle.

9. Turn on peristaltic injection pump to fill tube with liquid and purgeout all the air.

10. Adjust lighting for even illumination in the tank.

11. Set up camera in an appropriate area for viewing the entire Y-maze.

12. Fill Y-maze to the proper water level with 5 liters of sea watertaken from the shrimp holding system.

B. BETWEEN TRIALS USING THE SAME CHEMICAL

1. Rinse Y-maze and divider with warm tap water for approximately 30seconds.

2. Rinse with de-ionized water for 10 seconds.

3. Dry off outside of chamber and place on table.

4. Insert the appropriate tank dividers into the Y-maze.

5. Between the trials using the same chemical or concentration of thesame chemical, pump peristaltic injection pump tubing with at least 250mLs salt water.

6. Calibrate peristaltic injection pump to deliver at the predeterminedflow rate.

7. Place smaller tubing inflow and outflow ends into test chemicalbottle.

8. Turn on peristaltic injection pump to fill tube with liquid and purgeout all the air.

9. Adjust lighting for even illumination in the Y-maze.

10. Set up camera in an appropriate area for viewing the entire Y-maze.

11. Fill Y-maze to the proper water level with 5 liters of sea watertaken from the shrimp holding system.

2. TEST ORGANISM

Animals should be chosen from a previously isolated set of animals sorepetition does not occur within the test set for a particular chemicalconcentration. All test animals should be free of chitinolytic bacteriaif possible. Most importantly, animals should have all head and mouthappendages (antennules, antennae, maxillae, maxillipeds and walkinglegs).

1. Select an animal that appears to be calm (i.e., not repetitivelyhitting head into wall).

2. Net the animal carefully.

3. Place the animal in the appropriate position in the Y-maze (in thebase of the Y-maze).

4. If an animal jumps out of the tank or net at any time duringtransfer, return it to the holding chamber and choose another animal.

II. TEST PROCEDURE

1. PRETRIAL

1. When the animal is placed into the Y-maze, turn on the recirculationlines of pump and begin timing the acclimation period of 15 minutes.

2. Record the test date onto the data sheet.

3. When the acclimation period ends, turn on the camera to record animalmovements.

2. TRIAL

4. Immediately begin timing the run.

5. Insert feeding station with preweighed feed in place.

6. After 3 minutes, carefully remove any tank dividers.

7. Observe and record on a data sheet the placement and movements of theanimal. Also note any problems that occurred during the testing.

8. When the predetermined test time is over, turn off the peristalticinjection pump, camera and timer.

3. POST-TRIAL

9. Collect and place food in a drying oven to later record amoundingested by animal or dispersed into the water.

10. Remove Y-maze recirculation tubing and begin cleaning (see ApparatusPreparation above).

11. Pour the water from the Y-maze through a net into a waste bucket.The animal should now be in the net.

12. Place the animal into a group or individual holding chamber forlater testing.

13. Begin preparation of apparatus and test animal.

With respect to the Y-maze testing protocol, three tests were carriedout using the Y-maze of FIGS. 5A, 5C and 5E described in detail in theDetailed Description of the Drawings, infra.

Mixtures containing "natural extract" and glucose in saline solutionwere prepared and tested against a control which only had salinesolution.

In the following tables, "natural extract" is defined as a 1:1:1 weightratio of squid mantle:shrimp abdomen:crab claw. Sea water is usedcontaining 30 parts per 1,000 of FRITZ® salt.

EXAMPLE 1

    ______________________________________                                        1 ML NATURAL EXTRACT: 1 LITER SEA WATER: GLUCOSE                              (FINAL CONCENTRATION OF GLUCOSE 10.sup.-3 M)                                  Chemical        Figure                                                        Arm    Inserts  Reference                                                                              Reactions and Position in Tank                       ______________________________________                                        A      NONE     5A       animal (Paneaus vannamei) excited                                             but facing outlet                                    A      NONE     5A       animal jumped into A when gate                                                lifted                                               A      NONE     5A       animal jumped into A when gate                                                lifted                                               A      NONE     5A       went into arm A at 0:16                              B      CURVED   5E       facing back wall, slowly turned                                               and moved toward arms: turned to                                              side wall: turned to back wall:                                               turned to side wall: observed                                                 level 5 reactions                                    B      CURVED   5E       animal jumped when gate lifted                       B      CORRAL   5C       jumped to back when gate lifted:                                              at 4:40, animal came out and went                                             toward B: jumped into A                              B      CORRAL   5C       jumped into A when gate lifted                       B      CORRAL   5C       jumped when lifted gate, but                                                  stayed along back of corral:                                                  level 5 reaction                                     B      CORRAL   5C       animal never left position along                                              back of corral: could not see                                                 antennule or maxilliped movements                    ______________________________________                                    

EXAMPLE 2

    ______________________________________                                        1 ML NATURAL EXTRACT: 100 ML SEA WATER: GLUCOSE                               (AT A GLUCOSE CONCENTRATION OF 10.sup.-2 M)                                   Chemical        Figure                                                        Arm    Inserts  Reference                                                                              Reactions and Position in Tank                       ______________________________________                                        B      CORRAL   5C       animal jumped out of Y-maze                          B      CORRAL   5C       swimming around corral: back                                                  against outlet: into A: stayed in                                             A: extreme max and antennule                                                  movements                                            B      CORRAL   5C       animal "playing dead" in corral:                                              into back A by corral: into                                                   corral: into back of B: to B:                                                 long B-left wall: into corral:                                                extreme max and antennular                                                    movements                                            B      CORRAL   5C       into back A corner by corral                                                  facing back: across to B side                                                 wall to B back corner facing                                                  side: across to A wall: animal                                                became stressed and jumped into                                               B: remained in B                                     B      CORRAL   5C       into A: extreme antennule and                                                 maxilliped movements                                 B      CORRAL   5C       playing dead in corral until                                                  5:30: into back A corner: moved                                               to opening of A                                      A      CORRAL   5C       to A back corner facing A wall:                                               into A after a short stop: ran                                                only approximately 1 minute after                                             entry into A: extreme antennule                                               and maxilliped movements                             A      CORRAL   5C       no movements: stayed in corral                       B      CORRAL   5C       facing back of corral when                                                    started: backed out of corral,                                                then returned: animal hit head                                                into wall for a time: level 5                                                 reaction                                             A      CORRAL   5C       facing arms: turned to outlet end                                             at approximately 2:15: level 5                                                reactions                                            ______________________________________                                    

EXAMPLE 3

    ______________________________________                                        CONTROL (SOLELY SEA WATER)                                                    Chemical        Figure                                                        Arm    Inserts  Reference                                                                              Reactions and Position in Tank                       ______________________________________                                        NONE   CORRAL   5C       remained in corral: animal jumped                                             into A at approximately 7:00 when                                             adjusting camera: level 2 rxn                        NONE   CORRAL   5C       never left corral: level 3 or 4                                               reaction                                             NONE   CORRAL   5C       never left corral: level 4 or 5                                               reaction                                             NONE   CORRAL   5C       into A approximately 0:15 and                                                 remained in A                                        NONE   CORRAL   5C       jumped into A then back in corral                                             when gate lifted: slowly came out                                             into back corner of A: over to B                                              wall: slowly walked into B                           NONE   CORRAL   5C       jumped into A at approximately                                                4:00: jumped back into corral                                                 after 0:30: no outside stimuli                                                caused these reactions                               NONE   CORRAL   5C       never left corral                                    NONE   CORRAL   5C       active during acclimation:                                                    searched corral until went into A                                             at 9:00: jumped to A back at 9:55                    NONE   CORRAL   5C       went to A wall at approximately                                               4:00: to A back corner: slowly                                                turned and went into B after                                                  pausing and turning in front of A                    NONE   CORRAL   5C       animal jumped in air then back                                                into corral when gate lifted:                                                 never left corral                                    ______________________________________                                    

The following application of the feeding behavior:testing protocol wascarried out on Paneaus vannamei. Stock solutions were made up at aconcentration of 10⁻³ molar. 5 Ml of the stock solution is admixed with500 grams of feed causing 0.5 grams of feed to contain 1×10⁻⁸ moles oftest chemical. The volume in the aquarium where the feed testing istaking place is 150 liters. Accordingly, the average concentration ofthe test chemical is 7×10⁻¹² molar. When using 7 ml of test solutionrather than 5 ml, the final concentration will be 1×10⁻¹¹. The actualfeed is RANGEN® 35 containing 35% protein, 7% fat, fish meal (withoutsquid) fish oil, and minor amounts of nutrient vitamins and minerals.

The stock solution is diluted to 25 ml with sea water containing 30 pptof FRITZ® salt described, supra. 25 Ml of the stock solution is sprayedonto the 500 grams of feed. The water content is increased byapproximately 5%. The jar containing the solution is sealed and storedfor ten days at 0° C. Prior to use, the contents are shaken well.

The feed level is 0.5 grams of pellets per tank per feeding. Thefollowing diets J; P; A and C were used on Paneaus vannamei using thefeeding behavior testing protocol described, supra.

DIET J

7 Ml of 1×10⁻³ molar 2-methyl-3-(methyldithio)furan having thestructure: ##STR46## The solution is diluted to 25 ml with theartificial sea water containing 35 ppt of FRITZ® salt.

DIET P

N-acetyl-D-Glucosamine, an epimeric mixture of compounds having thestructures: ##STR47## Each of the solutions are 1×10⁻³ molar. The 21 mlis diluted to 25 ml with artificial sea water containing 30 ppt FRITZ®salt.

DIET A

2-Methyl-3-(methyldithio)furan having the structure: ##STR48##N-acetyl-D-Glucosamine, an epimeric mixture of compounds having thestructures: ##STR49## dimethyl sulfoxide having the structure: ##STR50##Osceola Brown Sugar produced by Osceola Farms Inc. of Pahokee, Fla.having a headspace analysis (trapped on TENAX® GC) as set forth in FIG.27, described in the Detailed Description of the Drawings, infra:3 ml;

a 50:50 mixture of skatole and indole, with the skatole having thestructure: ##STR51## and the indole having the structure: ##STR52##trimethyl amine oxide hydrate having the structure: ##STR53## the 24 ml(total) is diluted to 25 ml with artificial sea water.

DIET C (CONTROL DIET)

7 Ml of food grade ethyl alcohol (95%) is diluted to 25 ml withartificial sea water containing 30 ppt FRITZ® salt.

In grams, the overall growth of the Paneaus vannamei with each of thediets is set forth in the following table:

                  TABLE I                                                         ______________________________________                                        Diet       Overall Growth in Grams                                            ______________________________________                                        Diet C     1.43                                                               Diet A     1.22                                                               Diet J     1.58                                                               Diet P     1.36                                                               ______________________________________                                    

Thus, the most superior diet was Diet J, significantly better than theControl. Diet J contained the compound having the structure: ##STR54##

Results of the feeding studies using Diets C, A, J and P are summarizedin FIGS. 38A, 38B, 39, 40, 41, 42 and 43; described briefly and indetail, infra.

The following feeding model summarizes the use of the "ASIE" chemicalsof our invention as attractants, incitants, stimulants or excitants forthe members of the Penaeus genus of the Class Crustacea and sets forth asummary of the protocols of our invention: ##STR55##

It should be noted that the feeding "recipe" is as follows:

I. The following "alginate" mixture is prepared in the following manner:

1. Ingredients: A 2.5% aqueous solution of sodium alginate; and a 2.0%aqueous solution of sodium hexametaphosphate;

2. Apparatus: Two 750 ml beakers and two hot plates for use in heatingthe contents of the beakers; and

3. Procedure: Heat 500 ml of water to 80°-90° C. in one of the beakersusing one of the hot plates. Transfer 100 ml of the thus-heated water tothe second beaker. In the event that water-soluble attractants are beingused, the said water-soluble attractants are also added to the secondbeaker.

Solid sodium alginate is slowly added to the 100 ml of heated wateruntil the solution is at a level of 2.5% sodium alginate. Solid sodiumhexametaphosphate is then slowly added to the 100 ml alginate solutionuntil the sodium hexametaphosphate is at a level of 2%. The attractant,for example, the compound having the structure: ##STR56## (not solublein water) or Osceola Brown Sugar (soluble in water) is added as asolution (approximately 10-20 ml) in water (in the case of Osceola BrownSugar and other water-soluble materials) or in 95% aqueous food gradeethyl alcohol (in the case of the compound having the structure:##STR57##

The blending is to be continued for a period of 5 minutes in order toensure uniformity.

II. Feed Mixing Protocol

1. Grind the RANGEN® feed as fine as possible;

2. Place the feed into an industrial mixer and mix for a period of 30minutes;

3. Add the resulting "alginate" mixture from "I." and mix for a periodof 5 minutes;

4. If the resulting product has the proper amount of moisture, it willnot cake on the sides of the mixing vessel. Otherwise, it will adhere tothe sides of the mixing vessel as well as the mixing paddle. In thatevent, either additional water in an amount of 10-20 ml is required or ahigher speed (approximately 10% greater) for the paddle is required;

5. The final mix is to be extruded through a meat grinder using anappropriate die; and

6. In the event that the feed is still too dry, the surface of the feedwill flake and pellets will appear ragged. If the feed is too wet, theoperator will be unable to separate the strands as they extrude. Aproper amount of moisture will give rise to extrudates appearing assmooth tubes that are only slightly sticky.

The numerical results of the feed study for feed attractants A, C, J andP are set forth in the following Table IA:

                  TABLE IA                                                        ______________________________________                                        "FEED STUDY - % WEIGHT GAIN IN TEST DIETS"                                    BEG       END                                                                 AVE       AVE     % WEIGHT GAIN                                                                              allowance for rounding                         ______________________________________                                        A1     3.033  3.82    25.94790636                                             A2     2.889  3.579   23.88369678                                             A3     2.829  3.483   23.11770944                                             A AVE  2.917  3.634   24.58004799                                                                              24.31643753                                  C1     2.846  3.784   32:9585383                                              C2     2.721  3.411   25.35832415                                             C3     2.911  3.861   32.63483339                                             C AVE  2.826  3.679   30.18400566                                                                              30.31723195                                  J1     3.073  4.141   34.75431175                                             J2     2.996  4.058   35.44726302                                             J3     3.045  3.874   27.22495895                                             J AVE  3.038  4.029   32.62014483                                                                              32.47551124                                  P1     3.024  3.748   23.94179894                                             P2     2.855  3.84    34.50087566                                             P3     3.09   4.148   34.2394822                                              P AVE  2.99   3.904   30.56856187                                                                              30.89405227                                  ______________________________________                                    

Listed in the first column are each of the tank numbers for all of thediets. In the second column is the average weight in grams of that tankbefore the first day of the study. The row that says "A AVE", as well ascorresponding rows, to wit: "C AVE", "J AVE" and "P AVE", is the averagefor all three tanks for that specific diet. The third column is theaverage weight in grams for each tank and diet at the end of the study,that is, after six weeks. Then follows the percent weight gained by eachtank and the average percent gained for the corresponding diet. Theaverage of the combined percentages for the three tanks is thencalculated and appears in the final column with appropriate allowancefor "rounding" of the numerical data. Bold numbers are those used inFIGS. 38A and 38B comparing the percent growth for each diet. FIGS. 38Aand 38B are described briefly and in detail, infra, in the "BRIEFDESCRIPTION OF THE DRAWINGS" section and in the "DETAILED DESCRIPTION OFTHE DRAWINGS" section, infra.

Another aspect of our invention is the combination of the use of thestatic tank apparatus and procedure with the flow-through tank apparatusand procedure.

Another aspect of our invention is the use of the combination of thestatic tank apparatus, the flow-through apparatus and the Y-mazeapparatus and the procedures covering same.

With respect to the Procambrus clarkii species work, the volume of waterfor containing a member of the Procambrus clarkii species of the ClassCrustacea (against which a chemical is to be tested) is also a dilutesaline solution containing 30 parts per thousand of a FRITZ® Super Saltsolution described, supra.

Also with reference to the Procambrus clarkii species of the ClassCrustacea, the amount of attracting, stimulating, exciting or incitingchemical such as the trimethyl amine oxide hydrate having the structure:##STR58## in the salt solution may vary from about 10⁻⁴ gram moles perliter up to about 10⁻²¹ gram moles per liter; and the most preferableconcentration range varies from about 10⁻⁶ gram moles per liter down toabout 10⁻¹² gram moles per liter in the saline solution.

We have measured responses of Procambrus clarkii on a scale of 0-6 andthe measurement of such responses to various concentrations of variousattracting, exciting, stimulating and inciting chemicals is set forth indetail in the "DETAILED DESCRIPTION OF THE DRAWINGS" section of theinstant Application, infra.

The following results were obtained in the static system of FIG. 7B(described, infra) using the following parameters: all crawfish(Procambrus clarkii species of the Class Crustacea) tested had an intactshell and visible head appendages. All were acclimated in the staticchamber for 10 minutes and then run for 5 minutes. All positions andmovements within the tank were recorded, along with the excitatoryresponses they exhibited during that time. The following codes were usedto record their responses:

A=Antennule flicking;

AN₂ =Antennae movement;

M=Maxilliped movement;

MP=Mouthpart movement; and

D=Dactyl movement (in the characteristic "feeding" motion).

In the following groups of data, OBS means "Oseola Brown Sugar"manufactured by Osceola Farms Inc. of Pahokee, Fla.; NAG is an epimericmixture of compounds having the structures: ##STR59## Furthermore, suchterms as 10-6 stand for 10-6 gram moles per liter. The "SCORE" isdetermined as set forth in the DETAILED DESCRIPTION OF THE DRAWINGSsection, infra.

    ______________________________________                                        OBS 10.sup.-9 AVERAGE = 4.45                                                  CHEMICAL  CRAWFISH NO.  CODES       SCORE                                     ______________________________________                                        OBS 10.sup.-9                                                                           1             M, A, D     4                                         OBS 10.sup.-9                                                                           2             A, M, D     5                                         OBS 10.sup.-9                                                                           3             M, A        4                                         OBS 10.sup.-9                                                                           4             M, A, D     5                                         OBS 10.sup.-9                                                                           5             A, M        3                                         OBS 10.sup.-9                                                                           6             M, A        5                                         OBS 10.sup.-9                                                                           7             M, A, D     5                                         OBS 10.sup.-9                                                                           8             A, M          3.5                                     OBS 10.sup.-9                                                                           9             M, A        5                                         OBS 10.sup.-9                                                                           10                        X                                         OBS 10.sup.-9                                                                           11            A, M        5                                         OBS 10.sup.-9                                                                           12            A, M        4                                         OBS 10.sup.-9                                                                           13            A, AN.sub.2, M, D                                                                         5                                         OBS 10.sup.-12 AVERAGE = 3.67                                                 OBS 10.sup.-12                                                                          1             A, M        4                                         OBS 10.sup.-12                                                                          2             A, M        4                                         OBS 10.sup.-12                                                                          3             A, M        3                                         OBS 10.sup.-12                                                                          4             A, M        3                                         OBS 10.sup.-12                                                                          5             M, A        4                                         OBS 10.sup.-12                                                                          6             A, M          2.5                                     OBS 10.sup.-12                                                                          7             A, M, D      6*                                       OBS 10.sup.-12                                                                          8             A, M        3                                         OBS 10.sup.-12                                                                          9             M, A        3                                         OBS 10.sup.-12                                                                          10            A, M        4                                         OBS 10.sup.-12                                                                          11            A, M, D       3.5                                     OBS 10.sup.-12                                                                          12            A, M, MP    4                                         TMOH 10.sup.-6 AVERAGE = 3.54                                                 TMOH 10.sup.-6                                                                          1             M, A, MP, D 5                                         TMOH 10.sup.-6                                                                          2             A, M        3                                         TMOH 10.sup.-6                                                                          3             A, M, D     4                                         TMOH 10.sup.-6                                                                          4             M, A        5                                         TMOH 10.sup.-6                                                                          5             A, M        4                                         TMOH 10.sup.-6                                                                          6             A           1                                         TMOH 10.sup.-6                                                                          7             A, M        3                                         TMOH 10.sup.-6                                                                          8             A, M, AN.sub.2                                                                              3.5                                     TMOH 10.sup.-6                                                                          9                         0                                         TMOH 10.sup.-6                                                                          10            M, A, D, MP  6*                                       TMOH 10.sup.-6                                                                          11            A, M        4                                         TMOH 10.sup.-6                                                                          12            A, M        4                                         TMOH 10.sup.-12 AVERAGE = 3.42                                                TMOH 10.sup.-12                                                                         1             M, A, D     4                                         TMOH 10.sup.-12                                                                         2             A, M          2.5                                     TMOH 10.sup.-12                                                                         3             A, M, MP     6*                                       TMOH 10.sup.-12                                                                         4             A, M          2.5                                     TMOH 10.sup.-12                                                                         5             A, M          2.5                                     TMOH 10.sup.-12                                                                         6             A, M, D     4                                         TMOH 10.sup.-12                                                                         7             A, M          3.5                                     TMOH 10.sup.-12                                                                         8             M, AN.sub.2 3                                         TMOH 10.sup.-12                                                                         9             M, A        4                                         TMOH 10.sup.-12                                                                         10            A, M         6*                                       TMOH 10.sup.-12                                                                         11            M, AN.sub.2, A                                                                            3                                         TMOH 10.sup.-12                                                                         12                        0                                         BLANK AVERAGE = 2.38                                                          BLANK     1             A           2                                         BLANK     2             A, M          2.5                                     BLANK     3             A, AN.sub.2   1.5                                     BLANK     4             A, M        3                                         BLANK     5             M, AN.sub.2   2.5                                     BLANK     6             A, D        3                                         BLANK     7             A           2                                         BLANK     8             AN.sub.2, A 2                                         BLANK     9             M, A          3.5                                     BLANK     10            M, A          2.5                                     BLANK     11            A, AN.sub.2   2.5                                     BLANK     12            M, A          1.5                                     NAG 10.sup.-9 AVERAGE = 3.42                                                  NAG 10.sup.-9 M                                                                         1             M, AN.sub.2, MP, A                                                                        4                                         NAG 10.sup.-9 M                                                                         2             A, M        3                                         NAG 10.sup.-9 M                                                                         3             A, AN.sub.2, M                                                                            3                                         NAG 10.sup.-9 M                                                                         4             A, M, MP    5                                         NAG 10.sup.-9 M                                                                         5             M, AN.sub.2, A                                                                            3                                         NAG 10.sup.-9 M                                                                         6             A, AN.sub.2, M                                                                              3.5                                     NAG 10.sup.-9 M                                                                         7             A, M        3                                         NAG 10.sup.-9 M                                                                         8             D, AN.sub.2, A                                                                            4                                         NAG 10.sup.-9 M                                                                         9             A, M, D     3                                         NAG 10.sup.-9 M                                                                         10            AN.sub.2, A, MP                                                                           3                                         NAG 10.sup.-9 M                                                                         11            A, M, D       3.5                                     NAG 10.sup.-9 M                                                                         12            AN.sub.2, M, A                                                                            3                                         NAG 10.sup.-12 AVERAGE = 3.21                                                 NAG 10.sup.-12                                                                          1             A, D, M     3                                         NAG 10.sup.-12                                                                          2             A, M, AN.sub.2, D                                                                           3.5                                     NAG 10.sup.-12                                                                          3             A, MP, D, M 4                                         NAG 10.sup.-12                                                                          4             AN.sub.2, A, M                                                                              3.5                                     NAG 10.sup.-12                                                                          5             A, D, M     4                                         NAG 10.sup.-12                                                                          6             AN.sub.2, A, M                                                                              3.5                                     NAG 10.sup.-12                                                                          7             A           1                                         NAG 10.sup.-12                                                                          8             A, M        3                                         NAG 10.sup.-12                                                                          9             M, A        3                                         NAG 10.sup.-12                                                                          10            A, M, D     3                                         NAG 10.sup.-12                                                                          11            MP, M, AN.sub.2, A                                                                        4                                         NAG 10.sup.-12                                                                          12            A, M, D     3                                         NAG 10.sup.-18 AVERAGE = 3.17                                                 NAG 10.sup.-18                                                                          1             A, M          2.5                                     NAG 10.sup.-18                                                                          2             A, M, D     3                                         NAG 10.sup.-18                                                                          3             M, A, AN.sub.2                                                                            3                                         NAG 10.sup.-18                                                                          4             A, M        3                                         NAG 10.sup.-18                                                                          5             M, A, D     4                                         NAG 10.sup.-18                                                                          6             AN.sub.2, A   2.5                                     NAG 10.sup.-18                                                                          7             M, D, A       3.5                                     NAG 10.sup.-18                                                                          8             A, M          2.5                                     NAG 10.sup.-18                                                                          9             A, M        3                                         NAG 10.sup.-18                                                                          10            A, AN.sub.2 2                                         NAG 10.sup.-18                                                                          11            A, M, D       3.5                                     NAG 10.sup.-18                                                                          12            A, MP       3                                         ______________________________________                                                CHEMICAL                                                              CRAWFISH #                                                                              MOS 10.sup.-18                                                                          MOS 10.sup.-12                                                                          MOS 10.sup.-9                                                                         MOS 10.sup.-6                           ______________________________________                                        1         3         1         5       3.0                                     2         1.5       2         1.5     1                                       3         3         2         1       2.0                                     4         1         4         1.5     2.5                                     5         2         4         3       1                                       6         2.5       2.5       2.5     0                                       7         1.5       4.5       3       .5                                      8         2         1         4       1.5                                     9         3         2         4.5     2.5                                     10        2         1         3.5     .5                                      11        2.5       3         2.5     1.5                                     12        3         4.5       2.5     0                                       AVERAGE   2.25      2.63      2.88    1.33                                    MEDIAN    2.25      2.25      2.75    1.25                                    ______________________________________                                         *ANIMAL TOUCHED CHEM TUBE WITH MOUTHPARTS AND/OR CHELIPEDS               

The foregoing and additional data are set forth graphically in FIGS. 34,35, 36 and 37 described in the "BRIEF DESCRIPTION OF THE DRAWINGS" and"DETAILED DESCRIPTION DRAWINGS" sections, infra.

Specific aspects of the testing apparatus used in testing the abovechemicals as against the Procambrus clarkii species of the ClassCrustacea (crawfish) is set forth in the brief description of FIGS. 2and 3 and in the detailed description of FIGS. 2 and 3, infra, and isfurther set forth in copending Applications for U.S. Pat. Ser. No.08/279,181 filed on Jul. 22, 1994 and Ser. No. 08/413,440 filed on Mar.30, 1995, the specifications for which are incorporated herein byreference.

Response as a function of concentration of active chemical, e.g., TMOH,NAG and/or OBS taken alone or in combination is described via a numberof different mathematical models which are graphically set forth inFIGS. 6, 7 and 8 described, infra. These mathematical models are setforth as follows:

    R=5.0014- log.sub.10 C!.sup.2 (0.0084+0.5169e.sup.+0.45 log.sbsp.10.sup.C!)

(for NAG: mean of data);

    R=0.7874 -log.sub.10 C!.sup.-3/2 -0.0001 log.sub.10 C!.sup.2 +3.041

(for NAG: median of data);

    R=4.6574+0.0018 log.sub.10 C!.sup.3 +0.0322 log.sub.10 C!.sup.2 +0.0907 log.sub.10 C!

(for NAG: for mean of all data points);

    R=(-6×10.sup.-4) log.sub.10 C!e.sup.-0.2569 log.sbsp.10.sup.C! +0.0207 log.sub.10 C!+3.5607

(for TMOH: mean of data);

    R=-2.6824 sin h -0.1{log.sub.10 C}!-0.3680 log.sub.10 C!+3.1913

(for TMOH: for median of data points);

    R=(-0.0082) log.sub.10 C!.sup.2 e.sup.-0.0573 log.sbsp.10.sup.C! -0.0066 log.sub.10 C!.sup.2 +6.3210

(for OBS: mean of data points);

    R=(-1.5542×10.sup.4){log.sub.10 C}.sup.2 e.sup.+2.1941 log.sbsp.10.sup.C! -0.1613 log.sub.10 C!.sup.2 +11.2387

(for OBS: median of data points);

    R=0.1302 log.sub.10 C!.sup.2 e.sup.0.092 log.sbsp.10.sup.C! -0.3697 log.sub.10 C!-5.0549

(for OBS: mean of data points); and

    R=1.52.9674-0.732 log.sub.10 C!.sup.2 e.sup.0.0485 log.sbsp.10.sup.C +12.433 log.sub.10 C!

(for MOS: mean of data points).

In the foregoing equations, the term "R" is the "response" scored asdescribed, infra; and the term "C" is the concentration of activeingredient, e.g., OBS in the saline solution in gram moles per liter.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A is a schematic side elevation diagram of Penaeus vannamei,Pacific White Shrimp, farm-reared species, showing the locations ofvarious chemoreceptors as well as other appendages that are the basisfor measurement of excitation, incitation, attraction and stimulation ofthe Paneaus vannamei, a member of the Class Crustacea.

FIG. 1B is a schematic side elevation diagram of the Procambrus clarkii(crawfish) species of the Class Crustacea, showing the locations ofvarious chemoreceptors as well as other appendages that are the basisfor measurement of excitation, attraction and stimulation of theProcambrus clarkii (crawfish) species of the Class Crustacea.

FIG. 1C is a schematic bottom ("ventral") view of the Procambrus clarkii(crawfish) species of the Class Crustacea showing the locations ofvarious chemoreceptors as well as other appendages that are the basisfor measurement of excitation, attraction and stimulation of theProcambrus clarkii (crawfish) species of the Class Crustacea.

FIG. 1D is a schematic top ("dorsal") view of the Procambrus clarkii(crawfish) species of the Class Crustacea showing the locations ofvarious chemoreceptors as well as other appendages that are the basisfor measurement of excitation, attraction and stimulation of theProcambrus clarkii (crawfish) species of the Class Crustacea.

FIG. 2A is a schematic diagram of the static tank testing apparatususeful in the practice of our invention showing the interrelationship ofthe projection device 201 with the video camera or camcorder means 210of our invention.

FIG. 2B is a perspective view of the static tank 208 shown in the statictest apparatus of FIG. 2A.

FIG. 3 is a schematic cut-away side elevation view of apparatus showingthe use of the flow-through testing apparatus useful in the practice ofour invention.

FIG. 3A is a cut-away plan view of the flow-through tank apparatususeful in the practice of our invention taken in combination with thevideo camera/camcorder lens of the flow-through testing apparatus usefulin the practice of our invention shown in FIG. 3.

FIG. 3B is a cut-away side elevation view of the flow-through tankportion of the flow-through testing apparatus of FIG. 3 useful in thepractice of our invention.

FIG. 3C is an end view of the flow-through tank of the flow-throughtesting apparatus of FIG. 3 useful in the practice of our invention.

FIG. 4 is a cut-away side elevation view of the variable focus/focallength video camera or camcorder used in conjunction with the variableor constant intensity visible light and (i) the static tank testingapparatus useful in the practice of our invention; (ii) the flow-throughtesting apparatus useful in the practice of our invention or (iii) theY-maze apparatus useful in the practice of our invention.

FIG. 4A is a side elevation view of the support apparatus for the cameraused in conjunction with the apparatus of FIG. 4.

FIG. 4B is a front view of the support equipment for the camera of FIG.4.

FIG. 4C is a side elevation view of another embodiment of the apparatusused in conjunction with the camera of FIG. 4.

FIG. 4D is a front view of the support apparatus for the camera of FIG.4.

FIG. 4E is a top view of the support apparatus for the apparatus of FIG.4.

FIG. 4F is another embodiment of the support apparatus of the equipmentof FIG. 4.

FIG. 5A is the top view (cut-away) of the Y-maze testing apparatususeful in the practice of our invention without insertion of either a"corral" or a "curved wall" insert.

FIG. 5B is a cut-away side elevation view of the apparatus of FIG. 5A.

FIG. 5C is the top view of another embodiment of the Y-maze testingapparatus useful in the practice of our invention containing a "corral"section at the back of the base rectangular parallelepiped section ofthe Y-maze.

FIG. 5D is a cut-away side elevation view of the apparatus of FIG. 5C.

FIG. 5E is a top view (cut-away) of another embodiment of the Y-mazetesting apparatus useful in the practice of our invention containing a"curved wall" insert at the back of the base rectangular parallelepipedof the Y-maze apparatus.

FIG. 5F is a cut-away side elevation view of the Y-maze testingapparatus of FIG. 5E.

FIG. 5G is a schematic diagram illustrating the testing protocol usingthe Y-maze apparatus system useful in the practice of our invention.

FIG. 6A is a schematic diagram of Y-maze testing apparatus of the priorart.

FIG. 6B is a schematic block flow diagram showing production of OsceolaBrown Sugar manufactured by Osceola Farms, Inc. of Pahokee, Fla.

FIG. 7A is a perspective view of the static tank testing apparatus ofour invention shown in FIG. 2A, showing testing of Penaeus vannamei.

FIG. 7B is a perspective schematic diagram of the static tank testingapparatus useful in practicing our invention showing theinterrelationship of the projection device 201 with the video camera orcamcorder means 210 useful in practicing our invention; and showingtesting of Procambrus clarkii.

FIG. 7C is a perspective view in schematic form of the flow-throughtesting apparatus of FIG. 3 (without showing the camera blindcombination useful in the practice of our invention).

FIG. 7D is a schematic perspective view of apparatus showing the use ofthe flow-through testing apparatus useful in the practice of ourinvention; and showing testing of Procambrus clarkii.

FIG. 8 sets forth two graphs showing the response "R" versus: ##STR60##on the "Y" axis and the response ("R") on the "X" axis for the testingof the mixture of compounds having the structures: ##STR61## in thestatic tank testing apparatus.

FIG. 9 is a series of "Response as a function of concentration" graphs(parabolic and linear regression) showing on the "Y" axis:

     -log.sub.10 C!

and on the "X" axis the response "R" with "C" being in gram moles perliter for the materials:

(a) TALIN®, a mixture of Thaumatin I, Thaumatin II and Thaumatin B theliquid chromatograms of which are set forth in FIGS. 23 and 24 (TALIN®is a trademark of Tate and Lyle Limited of the United Kingdom);

(b) S-methyl methionine sulfonium chloride having the structure:##STR62## (c) D-Glucosamine, an epimeric mixture of compounds having thestructures: ##STR63## tested against Penaeus vannamei in a static tanktesting apparatus of FIGS. 2A and 7A.

FIG. 10 is a series of "Response as a function of concentration" graphs(parabolic and linear regression) of:

     -log.sub.10 C!

versus response "R" with:

     -log.sub.10 C!

being on the "Y" axis and "R" being on the "X" axis for the substances:

(a) trimethyl amine oxide hydrate having the structure: ##STR64## and(b) propiothetin (bromide) having the structure: ##STR65## in the statictank testing apparatus of FIGS. 2A and 7A as against Penaeus vannamei.

FIG. 11 is a series of "Response as a function of concentration" graphs(parabolic and linear regression) of:

     -log .sub.10 C!

versus response "R", with:

     -log.sub.10 C!

on the "Y" axis and "R" on the "X" axis for the substances:

(a) (R)(S)1-octen-3-ol having the structure: ##STR66## (a mixture ofisomers having the structures: ##STR67## and (b) guanidine having thestructure: ##STR68## in the static tank testing apparatus of FIGS. 2Aand 7A against the species Penaeus vannamei.

FIG. 12 is a "Response as a function of concentration" graph (linear)of:

     -log.sub.10 C!

on the "Y" axis versus response "R" on the "X" axis for TALIN® (amixture of Thaumatin I, Thaumatin II and Thaumatin B, the liquidchromatograms for which are set forth in FIGS. 23 and 24 described,infra) in the static tank testing apparatus for FIGS. 2A and 7A asagainst Penaeus vannamei. The concentration is in grams per liter.

FIG. 13 is a series of "Response as a function of concentration" graphs(parabolic and linear regression) of:

     -log.sub.10 C!

on the "Y" axis versus response "R" on the "X" axis for the substances:

(a) dimethyl sulfoxide having the structure: ##STR69## and (b) methionalhaving the structure: ##STR70## in the static tank test apparatus ofFIGS. 2A and 7A as against the species Penaeus vannamei.

FIG. 14 is a "Response as a function of concentration" pair of graphs(parabolic and linear regression) of:

     -log.sub.10 C!

on the "Y" axis versus response "R" on the "X" axis for the substance, a50:50 mole:mole mixture of skatole having the structure: ##STR71## andindole having the structure: ##STR72## in the static tank test apparatusof FIGS. 2A and 7A as against Penaeus vannamei.

FIG. 15 is a series of "Response as a function of concentration" graphs(parabolic and linear regression) of:

     -log.sub.10 C!

on the "Y" axis versus response "R" on the "X" axis for the substances:

(a) ammonium chloride;

(b) ammonia (aqueous); and

(c) acetic acid

using the static tank testing apparatus of FIGS. 2A and 7A as againstthe species Penaeus setiferus.

FIG. 16 is a series of "Response as a function of concentration" graphs(linear regression) of:

     -log.sub.10 C!

on the "Y" axis versus response "R" on the "X" axis for the substances:

(a) glycine;

(b) betaine having the structure: ##STR73## and (c) aspartate ion (assodium aspartate in solution)

in the static tank test apparatus of FIGS. 2A and 7A as against thespecies Penaeus setiferus.

FIG. 17 is a pair of "Response as a function of concentration" graphs(linear regression) of:

     -log.sub.10 C!

on the "Y" axis versus response "R" on the "X" axis for the substances:

(a) glucose; and

(b) taurine having the structure: ##STR74## in the static tank testingapparatus of FIGS. 2A and 7A as against Penaeus setiferus.

FIG. 18 is a schematic perspective diagram (bottom view) of thechemoreception test tank in the static tank testing apparatus of FIGS.2A and 7A.

FIG. 19 is a linear graph of flow rate (ml per minute) on the "Y" axisversus response "R" on the "X" axis for a mixture of glucose and"natural extract", a mixture of one part of equal portions of tissue ofcrab claw, squid mantle and shrimp abdomen in 1,000 parts of FRITZ®Super Salt Concentrate, "synthetic sea salt" solution, 30 parts perthousand, described in detail, supra. The ratio of "natural"extract:super salt solution:glucose solution being 1:1,000:1(weight:weight:weight). The graph is for the use of the solution in theflow-through apparatus of FIGS. 3 and 3B as against the species Penaeusvannamei.

FIG. 20 is a pair of "Response as a function of concentration" graphs(linear regression) of:

     -log.sub.10 C!

(on the "Y" axis) versus response "R" on the "X" axis for various flowrates at various concentrations of ammonium acetate using theflow-through apparatus of FIGS. 3 and 3B for the species Penaeusvannamei.

FIG. 21 is a "Response as a function of concentration" graph (linearregression) of:

     -log.sub.10 C!

(on the "Y" axis) versus response "R" on the "X" axis for the substanceN-acetyl-D-Glucosamine, an epimeric mixture of compounds having thestructures: ##STR75## in the flow-through chamber testing apparatus ofFIGS. 3 and 3B as against Penaeus vannamei.

FIG. 22 is a pair of "Response as a function of concentration" graphs(parabolic and linear regression) of:

     -log.sub.10 C!

on the "Y" axis versus response "R" on the "X" axis with "C" being ingram moles per liter for the substance, methional having the structure:##STR76## using the flow-through apparatus of FIGS. 3 and 3B as againstthe species Penaeus vannamei.

FIG. 23 is a liquid chromatogram profile for TALIN®, trademark of Tateand Lyle Limited of the United Kingdom, a mixture of Thaumatin I,Thaumatin II and Thaumatin B.

FIG. 24 is an HPLC (high pressure liquid chromatography) profile forTALIN®, the mixture of Thaumatin B, Thaumatin I and Thaumatin II,trademark of Tate and Lyle Limited of the United Kingdom.

FIG. 25 is a pair of "Response as a function of concentration" graphs(cubic and cubic regression) of:

     -log.sub.10 C!

on the "Y" axis versus response "R" on the "X" axis with "C" being ingram moles per liter for the substance, 2-methyl-3-(methyldithio)furanhaving the structure: ##STR77## in the flow-through chamber testingapparatus of FIGS. 3 and 3B as against Penaeus vannamei.

FIG. 26 is a series of "Response as a function of concentration" graphs(exponential, hyperbolic and linear regression) of:

     -log.sub.10 C!

on the "Y" axis versus response "R" on the "X" axis with "C" being ingram moles per liter for the substance Osceola Brown Sugar, manufacturedby Osceola Farms Inc. of Pahokee, Fla. (headspace analysis as trapped onTENAX® as set forth in FIG. 27) in the flow-through chamber testingapparatus of FIGS. 3 and 3B as against Penaeus vannamei.

FIG. 27 is a GC profile for the headspace analysis (trapped on TENAX®)for Osceola Brown Sugar, manufactured by Osceola Farms Inc. of Pahokee,Fla.

FIG. 28 is a "VENN" set diagram showing intersecting sets of stimulants,attractants, incitants and excitants for members of the species Penaeusgenus of the Class Crustacea and the shaded intersection for thosespecific substances which are each of incitants, stimulants, excitantsand attractants for members of the Penaeus genus of the Class Crustacea.

FIG. 29 is a pair of "Response as a function of concentration" graphs(hyperbolic or exponential and linear regression) of:

     -log.sub.10 C!

on the "Y" axis versus response "R" on the "X" axis with "C" being ingram moles per liter for the substance, N-acetyl-D-Glucosamine, anepimeric mixture of compounds having the structures: ##STR78## in theflow-through chamber testing apparatus of FIGS. 3 and 3B as againstPenaeus vannamei.

FIG. 30 is a pair of "Response as a function of concentration" graphs(mean and median) (exponential and linear regression) of:

     -log.sub.10 C!

on the "Y" axis versus response "R" on the "X" axis with "C" being ingram moles per liter for the substance, methional having the structure:##STR79##

FIG. 31 a pair of "Response as a function of concentration" graphs (meanand median) (parabolic) of:

     -log.sub.10 C!

on the "Y" axis versus response "R" on the "X" axis with "C" being ingram moles per liter for the substance, a 50:50 mixture of skatolehaving the structure: ##STR80## and indole having the structure:##STR81## in the flow-through chamber testing apparatus of FIGS. 3 and3B as against Penaeus vannamei.

FIG. 32 is a pair of "Response as a function of concentration" graphs(mean and median) (linear) of:

     -log.sub.10 C!

on the "Y" axis versus response "R" on the "X" axis with "C" being ingram moles per liter for the substance methionine having the structure:##STR82## in the flow-through chamber testing apparatus of FIGS. 3 and3B as against Penaeus vannamei.

FIG. 33 is a graph of "Response as a function of concentration" (linear)for the mean and median of data in the static tank apparatus of FIG. 2Afor yeast extract as against Penaeus vannamei.

FIG. 34 sets forth two "response as a function of concentration" graphssetting forth data for the response "R" versus: ##STR83## on the "Y"axis (and "C" being concentration in gram moles per liter) and theresponse "R" on the "X" axis for the testing of the mixture of compoundshaving the structures: ##STR84## in the static tank testing apparatus(of FIG. 7B) as against the Procambrus clarkii (crawfish) species of theClass Crustacea. The graph indicated by reference numeral 601' (a"gamma" function) is for the means of responses "R" versus:

     -log.sub.10 C!

The graph indicated by reference numeral 651' (a regression curve) isfor the medians of responses "R" versus:

     -log.sub.10 C!

FIG. 35 sets forth two "response as a function of concentration" graphs("gamma" function and hyperbolic function) of:

     -log.sub.10 C!

versus response "R", with:

     -log.sub.10 C!

being on the "Y" axis (and "C" being concentration in gram moles perliter) and "R" being on the "X" axis for the testing of the substancetrimethyl amine oxide hydrate having the structure: ##STR85## in thestatic tank testing apparatus of FIG. 7B against Procambrus clarkii. Thegraph indicated by reference numeral 701' (a "gamma" function) is forthe means of responses "R" versus:

     -log.sub.10 C!

The graph indicated by reference numeral 751' (a "hyperbolic" function)is for the medians of responses "R" versus:

     -log.sub.10 C!

FIG. 36 sets forth "response as a function of concentration" graphs(means of response indicated by reference numeral 801' (a "gamma"function) and medians of response as indicated by reference numeral 851'(also a "gamma" function)) of:

     -log.sub.10 C!

on the "Y" axis versus response "R" on the "X" axis with "C" being ingram moles per liter for the substance Osceola Brown Sugar, manufacturedby Osceola Farms Inc. of Pahokee, Fla. (headspace analysis as trapped onTENAX® as set forth in FIG. 27) in the static tank testing apparatus ofFIG. 7B as against the Procambrus clarkii species of the ClassCrustacea.

FIG. 37 sets forth two "response as a function of concentration" graphssetting forth data for the response "R" versus: ##STR86## on the "Y"axis (and "C" being concentration in gram moles per liter) and theresponse "R" on the "X" axis for the testing of the compound, MOS,having the structure: ##STR87## in the static tank testing apparatus of(FIG. 7B) as against the Procambrus clarkii (crawfish) species of theClass Crustacea. The graph indicated by reference numeral 3710 (a"gamma" function) is for the means of responses "R" versus:

     log.sub.10 C!

The graph indicated by reference numeral 3700 (a "gamma" function) isfor the median of responses "R" versus:

     -log.sub.10 C!.

FIG. 38A is a series of bar graphs summarizing the feeding studies usingDiets A, C, J and P for members of the Penaeus genus of the ClassCrustacea with the given diets being on the "X" axis and and the percentweight gain for members of the Penaeus genus of the Class Crustacea onthe "Y" axis.

FIG. 38B is another series of bar graphs for feeding Diets A, C, J and Pfor members of the Penaeus genus of the Class Crustacea showing averagegrowth over a period of six weeks in grams on the "Y" axis versus theparticular diet (A, C, J and P) on the "X" axis.

FIG. 38C is another series of bar graphs showing, for feeding Diets A,C, J and P for members of the Penaeus genus of the Class Crustacea, thetotal number of "hits" over a period of six weeks (on the "Y" axis)versus the particular diet on the "X" axis (that is, Diets A, C, J andP). The number of "hits" is the number of times that the member of thePenaeus genus of the Class Crustacea touches the feeding compositionduring the study period.

FIG. 39 is a graph showing on the "X" axis the weight gain (ΔW) formembers of the Penaeus genus of the Class Crustacea versus the number of"hits" (in units of hundreds of hits, H) on the "Y" axis. Two graphs areshown, graph 3900, an "exact fit" curve defined according to theequation: ##EQU19## and a linear regression line, indicated by referencenumeral 3910 defined by equation:

    H=66.67(ΔW)-44.33.

FIG. 40 is a linear regression line for the differences for each ofDiets J, P and A with respect to the control Diet C; with differences ingrowth of the members of the Penaeus genus of the Class Crustacea (ingrams) on the "X" axis and differences in number of "hits", ΔH, on the"Y" axis, the linear regression line being shown by reference numeral4000. The symbol for the differences in growth is given by the term:Δ(ΔW). The linear regression line is defined according to the equation:

    ΔH=5333.33 (Δ ΔW!)-180.

FIG. 41 is a series of graphs showing the difference in the weightgrowth for Diet J and for Diet P for members of the Penaeus genus of theClass Crustacea (Penaeus aztecus and Penaeus vannamei) (on the "X" axis)versus the percent of "active" on the "Y" axis; "active" being indicatedby the letter "A" and the "active" being either Diet J, the compoundhaving the structure: ##STR88## or Diet P, a mixture of the compounds:(i) N-acetyl-D-Glucosamine;

(ii) dimethyl sulfoxide; and

(iii) methional

in equal proportions. The activity of Diet A in the series of graphs isset forth as a result of the fact that Diet A contains lesser quantitiesof both the compound having the structure: ##STR89## as well as themixture of compounds: (i) N-acetyl-D-Glucosamine;

(ii) dimethyl sulfoxide; and

(iii) methional.

The graphs for Diet P are indicated by reference numerals 4100 and 4110.The graphs for Diet J are indicated by reference numeral 4120 and 4130.Graph 4100 (for Penaeus vannamei) is defined by the equations: ##EQU20##

The graph indicated by reference numeral 4110 (for Penaeus aztecus) isdefined according to the equation: ##EQU21##

The graph defined according to reference numeral 4120 (for Penaeusvannamei; Diet J) is defined according to the equation: ##EQU22##

The graph indicated by reference numeral 4130 for Penaeus aztecus (usingDiet J) is defined according to the equation: ##EQU23##

The feeding study results are also summarized in FIGS. 42 and 43.

FIG. 42 is a three-dimensional graph of difference in the weight gain(from control) on the "Y" axis versus log₁₀ (of the change in the numberof "hits") (on the "X" axis) versus log₁₀ A (log₁₀ percent active!) onthe "Z" axis for Diet P. The graph, indicated by reference numeral 4200,is defined according to the equations: ##EQU24##

FIG. 43 is a graph using the same coordinates as FIG. 42, except that itis for Diet J rather than Diet P. The graph indicated by referencenumeral 4300 is defined according to the equation:

    3.647 log.sub.10 (ΔH)!-1.636(Δ ΔW!)+0.345 log.sub.10 A!=10.

FIG. 44 is a pair of "response as a function of concentration" graphs(straight line) of:

     -log.sub.10 C!

on the "Y" axis versus response "R" on the "X" axis with "C" being ingram moles per liter for the substance "OXYCYCLOTHIONE-030™", having thestructure: ##STR90## using the static tank apparatus of FIGS. 2, 2A and7A as against the species Penaeus vannamei. The graph indicated byreference numeral 4400 is for the median data points. The graphindicated by reference numeral 4410 is for the mean of the data points.The graph indicated by reference numeral 4400 is defined according tothe equation:

    R=8.5+0.5 log.sub.10 C!.

The graph indicated by reference numeral 4410 is defined according tothe equation:

    R=7.3792+0.4167 log.sub.10 C!.

DETAILED DESCRIPTION OF THE DRAWINGS

FIG. 1A shows a cut-away schematic diagram of Penaeus vannamei species.The Penaeus vannamei is shown by reference numeral 10. Referencenumerals 11a and 11b refer to the antennal flagellum. Reference numeral12 refers to the antennular medial flagellum. Reference numeral 13refers to the lateral antennular flagellum. Reference numeral 14 refersto the antennal scale. Reference numeral 15a refers to a dactyl ofmaxilliped, the maxillipedes being indicated by reference numerals 15c,15d and 15b. Reference numeral 16 refers to the mandible, maxillule andmaxilla. Reference numerals 17e, 17f, 17g, 17h and 17i refer topereiopods. Reference numerals 17a and 17c refer to dactyls ofpereiopods. Reference numerals 17b and 17d refer to the merus ofpereiopods. Reference numerals 19a, 19b, 19c, 19d and 19e refer to thepleopods. Reference numerals 20a and 20b refer to uropods. Referencenumeral 18 refers to the branchial chamber of the Penaeus vannamei.

In the static tank means of FIGS. 2A and 7A, the following is theestablished grade protocol with respect to the Penaeus vannamei of FIG.1A as shown by reference numeral 10:

                  TABLE II                                                        ______________________________________                                        Value      Description and Figure Reference Numeral                           ______________________________________                                        X          invalid run                                                        (not counted and                                                              not made part of                                                              calculation)                                                                  0          no apparent reaction                                                          sporadic maxilliped beating (reference                                        numerals 15a, 15b and 15c); no antennular                                     activity (reference numeral 13)                                    2          regular maxilliped beating (reference numerals                                15a, 15b and 15c); no antennular activity                                     (reference numeral 13)                                             3          regular maxilliped beating (reference numerals                                15a, 15b and lSc); sporadic maxilliped beating                                (reference numerals 15a, 15b and 15c)                              4          continuous maxilliped beating (reference                                      numerals 15a, 15b and 15c); sporadic                                          maxilliped beating (reference numerals 15a,                                   15b and 15c)                                                       5          continuous maxilliped beating (reference                                      numerals 15a, 15b and 15c); extreme antennular                                activity (reference numeral 13)                                    ______________________________________                                    

With regard to the order of motion priorities, the following is thelikelihood of motion priorities in investigating the Penaeus vannameishown by reference numeral 10 on FIG. 1A:

                  TABLE III                                                       ______________________________________                                        Priority    Reference Numerals                                                ______________________________________                                        1           15 and 16                                                         2           12 and 13                                                         3           17                                                                ______________________________________                                    

With respect to prioritized order of importance of motion, the followingrefers to the Penaeus vannamei shown by reference numeral 10 on FIG. 1A:

                  TABLE IV                                                        ______________________________________                                        Priority    Reference Numerals                                                ______________________________________                                        1           17                                                                2           15                                                                3           16                                                                4           13                                                                5           12                                                                ______________________________________                                    

Referring to the schematic diagrams of the Procambrus clarkii species,FIGS. 1B, 1C and 1D, the Procambrus clarkii species is shown byreference numeral 100. FIG. 1B shows the scheme of internal organizationof the Procambrus clarkii species. FIG. 1C shows a ventral view of theProcambrus clarkii species. FIG. 1D shows a dorsal view of theProcambrus clarkii species. Reference numeral 1' refers to the compoundeye; reference numeral 2' refers to the rostrum; reference numeral 3'refers to the carapace of cephalothorax; reference numeral 4' refers tothe cervical groove; reference numeral 5' refers to one of severalantennules; reference numeral 6' refers to one of two antennae;reference numeral 7' refers to one of several maxillipeds; referencenumeral 7a' refers to the one of several dactyls; reference numeral 8'refers to one of several pereiopods (chelipeds); reference numeral 9'refers to one of several pleopods; reference numeral 10' refers to oneof several abdominal somites; reference numeral 11' refers to thetelson; and reference numeral 12' refers to one of several uropods.

Reference numerals 13', 14', 15', 16', 17' and 18' refer to thedigestive system of the Procambrus clarkii species of the ClassCrustacea. Reference numeral 13' refers to the mouth of the Procambrusclarkii species. Reference numeral 14' refers to the esophagus;reference numeral 15' refers to the cardiac stomach; reference numeral16' refers to the pyloric stomach; reference numeral 17' refers to theintestine; and reference numeral 18' refers to the digestive gland.

Reference numerals 19', 20' and 21' refer to the excretory system of theProcambrus clarkii species of the Class Crustacea. Reference numeral 19'refers to the green gland; reference numeral 20' refers to the bladder;and reference numeral 21' refers to the excretory pore.

Reference numerals 22' and 23' refer to the reproductive system of theProcambrus clarkii species of the Class Crustacea Reference numeral 22'refers to the testis; reference numeral 23' refers to the sperm duct.

Reference numerals 24', 25', 26', 27', 28', 29', 30', 31' and 32' referto the circulatory system of the Procambrus clarkii species of the ClassCrustacea. Reference numeral 24' refers to the heart; reference numeral25' refers to the ostium; reference numeral 26' refers to the ophthalmicartery anterior aorta; reference numeral 27' refers to the antennaryartery; reference numeral 28' refers to the dorsal abdominal artery;reference numeral 29' refers to the sternal artery; reference numeral30' refers to the segmental artery; reference numeral 31' refers to theventral abdominal artery; and reference numeral 32' refers to theventral thoracic artery.

With respect to the nervous system of the Procambrus clarkii species ofthe Class Crustacea, reference numeral 33' refers to the brain; andreference numeral 34' refers to the ventral nerve cord.

In the static tank means of FIG. 7B, the following is the establishedgrade protocol with respect to the Procambrus clarkii species of theClass Crustacea of FIGS. 1B, 1C and 1D as shown by reference numeral100:

                  TABLE V                                                         ______________________________________                                        Value        Description and Figure Reference Numeral                         ______________________________________                                        X            invalid run                                                      (not counted                                                                  and not made                                                                  part of                                                                       calculation)                                                                  0            no apparent reaction                                             1            sporadic maxillary movement (reference                                        numeral 7'); no antennule activity                                            (reference numeral 5'); and no dactyl                                         movement (reference numeral 7a')                                 2            regular maxillary movement (reference                                         numeral 7'); no antennule activity                                            (reference numeral 5'); and no dactyl                                         movement (reference numeral 7a')                                 3            regular maxillary movement (reference                                         numeral 7'); sporadic antenule activity                                       (reference numeral 5'); and little or no                                      dactyl movement (reference numeral 7a')                          4            continuous maxillary movement (reference                                      numeral 7'); sporadic antennule activity                                      (reference numeral 5'); and sporadic dactyl                                   movement (reference numeral 7a')                                 5            continuous maxillary movement (reference                                      numeral 7'); extreme antennule activity                                       (reference numeral 5'); and regular dactyl                                    movement (reference numeral 7a')                                 6            continuous maxillary movement (reference                                      numeral 7'); extreme antennule activity                                       (reference numeral 5'); regular dactyl                                        movement (reference numeral 7a'); and                                         contact between mouthparts and/or chelipeds                                   (reference numerai 8') with chem tube.                           ______________________________________                                    

In each of the above descriptions of Table V, the dactyl movement(reference numeral 7a') is defined as the characteristic "picking andfeeding" movement described above with the dactyls of the pereipods(reference numeral 7a') are touched to the bottom and then drawn to themouth.

The Procambrus clarkii species (crawfish) of the Class Crustacea possessthe characteristic set of crustacean head appendages which include ashort pair of antennules (reference numeral 5') and a longer pair ofantennae (reference numeral 6' of FIG. 1C). Their third maxilliped(reference numeral 7') is enlarged and in close proximity to the mouth(reference numeral 13'). The Procambrus clarkii species of the ClassCrustacea has a highly modified, enlarged first pereiopod (referencenumeral 8') with a large chelate dactyl called a cheliped.

Thus, for example, in 12 runs of 9 different animals (the Procambrusclarkii species of the Class Crustacea) using a comparison of a freshwater blank to Osceola Brown Sugar, the following responses were notedas positive:

(1) flicking of antennules (reference numeral 5' of FIG. 1C). The speedof flicking increased as the chemical was introduced, while the blankruns showed no increase in antennule flicking;

(2) waving of antennules (reference numeral 5') in a dorso-ventraldirection;

(3) maxillary movement (reference numeral 7') constant and increasedspeed during chemical runs;

(4) dacytl movement (reference numeral 7a') in a "grabbing and feeding"manner. Dacytls "pick" at bottom and are then brought to the mouth(reference numeral 13') area. Frequency increased as chemicalconcentration in the static chamber of FIG. 7B increased and no dactylmovement of this type was seen in the blank runs; and

(5) in all chemical runs, animals eventually oriented themselves towardthe chemical tube and were seen to move the tube to their mouthpartswith the pereiopods (reference numeral 8') and third maxillipeds(reference numeral 7'). When the pump was turned off, stopping thechemical flow, the tube was released after approximately one minute (thetube is indicated in FIG. 7B as tube 204b).

Thus referring to the apparatus of FIG. 7B, the static testing system,the static tank system is shown by reference numeral 200. Referencenumeral 201 refers to the video monitor. Reference numeral 203 is thevessel holding testing solution 202. Testing solution 202 is pumpedthrough line 204a using peristaltic multihead pump 206 controlled bydevice 205, pumping solution 202 into tank 208. Tank 208 contains liquid212 in which the Procambrus clarkii species of the Class Crustacea is atrest on the bottom of the tank, which bottom is indicated by referencenumeral 214 which may be entirely composed of a clear plastic or glassso that the video camera or camcorder 210 may be focused on the movementof the member of the Procambrus clarkii species of the Class Crustacea100 utilizing the fluorescent light-generating device 211. Tank 208 ismounted on stand 209 which surrounds the video camera or camcorder 210directed towards the tank 208. The surface of the liquid 212 is shown byreference numeral 213.

An example of the light apparatus 211 is a Flexo Heavy-Duty AdjustableLamp made by Art Specialty Company holding two 18" fluorescent tubes.The tubes are manufactured by the General Electric Company ofSchenectady, N.Y. The following specifications for the tubes used are anexample of what can be used in the operation of the apparatus of FIG.7B:

GE Catalog No. F15T8/CW;

18" Cool White fluorescent tube;

15 Watt;

Rated life of 7,500 hours;

Initial lumens=825;

Mean lumens=725;

Kelvin temperature=4,150; and

CRI rating of 62.

The peristaltic pumps utilizable are those, for example, identified asMASTERFLEXφ L/S manufactured by the Cole-Parmer Instrument Company of7425 North Oak Park Avenue, Chicago, Ill. 60648 (MASTERFLEX® being atrademark owned by Cole-Parmer Instrument Company). The pump head is astandard pump head. An example of the MASTERFLEXφ tubing utilized withthe MASTERFLEX® tubing pump (peristaltic pump) is C-FLEX® 06424(trademark of Cole-Parmer Instrument Company), astyrene-ethylene-butylene modified block copolymer.

Referring to FIG. 7D, the flow-through vessel testing apparatus whichcan also be used in testing substances with respect to attraction of theProcambrus clarkii species of the Class Crustacea, fluid to be tested302 is contained in container 303 and pumped through tube 304a usingpump 306b and then through tube 304b into location 325 and then intovessel 350. Vessel 350 contains the Procambrus clarkii species of theClass Crustacea 10' and 10" located in the flowing liquid 312.Meanwhile, the liquid 312 is circulating by means of pump 306a throughline 324a and then through 324b into tube 326a and fitting 331a where itjoins with the feeding fluid (test material) at 325. The combinedliquids having ever-increasing concentration of material from vessel 303travels through holding vessel 350 into exit tube 331b past fitting 329into tube 326b where, again, it is recirculated. The exit portion of theflow-through tank is 351b and the entrance portion is 351a.

The Procambrus clarkii animal is placed in section 351b to acclimatewith only saline water 312 circulating. The experiment is started byadding liquid (302) to the liquid 312 and removing the barrier 330. Themotion of the Procambrus clarkii animal 100 from position 10" to 10' ismonitored by video camera or camcorder 310 and displayed on displaydevice 301.

Meanwhile, the motions of the member of the Procambrus clarkii speciesof the Class Crustacea 10' and 10" is recorded using video camera orcamcorder 310 shown on monitor 301. Simultaneously, light source 311directs light into flow-through vessel 350. The side of the flow-throughvessel has a clear plate through which camera 310 has a direction ofvision. Screen 330 is held in place at 335. Screen 330 divides theflow-through tank between sections 351a, the entrance section and 351b,the exit section.

Additional testing fluid enters through tube 304b and enters the elbow,mixing with fluid from 326a at 331a. Fitting 326a is threaded into elbow325 at 329. Fluid exits entering elbow 331b. Fitting 326b is threadedinto the elbow at 329.

Referring to the apparatus of FIG. 2A, the static tank testing system,the static tank system is shown by reference numeral 200. Referencenumeral 201 refers to the video monitor. Reference numeral 203 is thevessel holding testing solution 202. Testing solution 202 is pumpedthrough line 204a using peristaltic multihead pump 206 controlled bydevice 205 pumping solution 202 into tank 208. Tank 208 contains liquid212 in which Penaeus vannamei or one of the members of the Penaeus genusof the Class Crustacea is at rest on the bottom of the tank which bottomis indicated by reference numeral 214 which may be entirely composed ofa clear plastic or glass so that video camera or camcorder 210 may befocused on the movement of the member of the Penaeus genus of the ClassCrustacea 10 utilizing the fluorescent light generating device 211. Tank208 is mounted on stand 209 which surrounds the video camera orcamcorder 210 directed towards the tank 208. The surface of the liquid212 is shown by reference numeral 213. FIG. 7A is a perspective view ofthe apparatus of FIG. 2A with each of the reference numerals of FIG. 2Arepeated.

An example of the light apparatus 211 is a Flexo Heavy-Duty AdjustableLamp made by Art Specialty Company holding two 18" fluorescent tubes.The tubes are manufactured by the General Electric Company ofSchenectady, N.Y. The following specifications for the tubes used are anexample of what can be used in the operation of the apparatus of FIGS.2A and 7A:

GE Catalog No. F15T8/CW;

18" Cool White fluorescent tube;

15 Watt;

Rated life of 7,500 hours;

Initial lumens=825;

Mean lumens=725;

Kelvin temperature=4,150; and

CRI rating of 62.

The peristaltic pumps utilizable are those, for example, identified asMASTERFLEX® L/S manufactured by the Cole-Parmer Instrument Company of7425 North Oak Park Avenue, Chicago, Ill. 60648 (MASTERFLEX® being atrademark owned by Cole-Parmer Instrument Company). The pump head is astandard pump head. An example of the MASTERFLEX® tubing utilized withthe MASTERFLEX® tubing pump (peristaltic pump) is C-FLEX® 06424(trademark of Cole-Parmer Instrument Company), astyrene-ethylene-butylene modified block copolymer.

Referring to FIG. 3, the flow-through vessel testing apparatus, fluid tobe tested 302 is contained in container 303 and pumped through tube 304ausing pump 306b and then through tube 304b into location 325 and theninto vessel 350. Vessel 350 contains the member of the Penaeus genus ofthe Class Crustacea 10' and 10" located in the flowing liquid 312.Meanwhile, the liquid 312 is circulating by means of pump 306a throughline 324a and then through 324b into tube 326a and fitting 331a where itjoins with the feeding fluid (test material) at 325. The combinedliquids having ever-increasing concentration of material from vessel 303travels through holding vessel 350 into exit tube 331b past fitting 329into tube 326b where again it is recirculated. The exit portion of theflow-through tank is 351b and the entrance portion is 351a.

The Penaeus vannamei is placed in section 351b to acclimate with onlysaline water 312 circulating. The experiment is started by adding liquid(302) to the liquid 312 and removing the barrier 330. The motion ofPenaeus vannamei 10 from position 10" to 10' is monitored by 310 anddisplayed on 301.

Meanwhile, the motions of the member of the Penaeus genus of the ClassCrustacea 10' and 10" is recorded using video camera or camcorder 310shown on monitor 301. Simultaneously, light source 311 directs lightinto flow-through vessel 350. The side of the flow-through vessel has aclear plate through which camera 310 has a direction of vision. Screen330 is held in place at 335. Screen 330 divides the flow-through tankbetween sections 351a, the entrance section and 351b, the exit section.As shown in FIGS. 3A, 3B and 3C, fluid entering the flow-through holdingtank 350 enters from tube 352a and exits from tube 352b. Additionaltesting fluid enters through tube 304b and enters the elbow mixing withfluid from 326a at 331a. Fitting 326a is threaded into elbow 325 at 329.Fluid exits at 352b entering elbow 331b. Fitting 326b is threaded intothe elbow at 329.

Referring to FIG. 4, FIG. 4 shows a side elevation view partiallycut-away of apparatus showing in detail the variable focus/focal lengthvideo camera or camcorder, camera platform and shield used with thelight in conjunction with the test chamber. Chamber 408 could be aflow-through chamber containing fluid 412 therein. Light source 411emits visible wavelength radiation into the tank 408 and into the fluid412 simultaneously with the action of video camera 410 strapped toplatform 414 with bungee cord 421. Video camera 410 is mounted on aheight adjuster 452 having a movable height adjustment screw 413 andimbedded ball bearings 430 at a solid surface indicated by 415. Thecamera is mounted so that the lens, lens shield 470 and the liquid 412are on a direct visual line as a result of an orifice being insupporting frame 419 and another orifice indicated by reference numeral418 being in shield 417. Supporting frame 419 and the camera lens areshielded by cloth 420 which prevents the member of the Penaeus genus ofthe Class Crustacea 10 from being distracted by the movement of thevideo camera. Thus, camera 410 may be adjusted laterally and vertically,the vertical angle adjustment coming through the use of adjustment screw413 and the lateral travel coming through the travel using bearings 430on the solid travel surface or lamina 415.

FIG. 4A is a detailed section of the shield 417 having attached theretoa supporting frame guide 422 at an angle preferably of 45° to the shield417. 418 indicates the orifice mentioned in the detailed description ofFIG. 4. FIG. 4B is a front view of the frame of FIG. 4A showing theorifice 418 and the supporting frame guide 422 and the vertical part ofthe frame 417. FIG. 4C is another side elevation view of the cameraplatform showing via hidden lines the adjustable camera mount platform452 and the hinge to which the adjustable mount 452 is connected to theframe, the hinge being indicated by reference numeral 450.

FIG. 4D is the front view of the camera platform looking in thedirection of the camera from the flow-through tank. The cloth cover 420is connected via a frame 419 to a main cover 417. The orifice in thecloth 420 is indicated by reference numeral 418a, through which isplaced lens shield 470 of video camera or camcorder 410.

FIG. 4E is a top view of the camera platform showing the adjustableplatform 452 and the adjustable screw 413 with hinges 450 and showingthe location of the inner frame 419.

FIG. 4F is a cut-away side elevation view showing, in detail, theadjustable platform 452 and the adjustable screw 413 with hinges 450 andshowing the location of inner frame 419.

FIG. 5A is a top cut-away view of a Y-maze apparatus for testingchemo-attractancy of a substance. The Y-maze apparatus is indicated ingeneral by reference numeral 500. The member of the Penaeus genus of theClass Crustacea 10 may or may not be attracted to an attractant or itmay or may not be excited by an excitant. In any event, the feed lineinflow fittings 560 are located at the end of the Y-maze 509a and 509b.

Thus, the test section of the Y-maze is indicated by 509a and thecontrol section is indicated by 509b or vice versa (these two sectionscan be reversed by changing the chemical feed lines as shown in FIG. 5Gand discussed in detail, infra). Both sections join at 540 with ajoining wall 543.

The "control section" and the "testing substance input section" (alsoindicated by the letters "A" and "B" which can be interchanged) are atan angle "alpha" from one another, alpha being 5°-45°, preferably 10°.These sections as stated, supra, are interchangeable. The vertex of theangle is at reference numeral 540. An optional feeding station alsoexists in the Y-maze test section and is indicated by reference numeral596 and in the control section at 597. Baffle plates 501a, 501b, 501cand 501d (also called "flow screens") are located close to the end 509aand 509b close to the inflow fitting 560 coming from the feed fluidperistaltic pump. The testing section is indicated overall by referencenumeral 520 or 521 and the control section is indicated overall byreference numeral 520 or 521 since the two sections can be reversed bychanging the chemical feed lines as shown in the detailed description ofFIG. 5G, infra. The feeding section and the control section areseparated by a removable screen 503 from the main section of the Y-maze522 in which the member of the Penaeus genus of the Class Crustacea 10is located. The sides of the main section 522 are indicated by referencenumerals 508a and 508b. The back end of the Y-maze 522 which is in theshape of a rectangular parallelepiped contains two drain lines 562a and562b and a flush port 561, the back section being indicated by referencenumeral 526. A baffle between the back section 526 and the main sectionof the Y-maze 522 is indicated by reference numeral 590. The flow ofliquid travels through portal 570 underneath perforated wall 590 andaround baffle 524 to the drain. The fluid level is also shown in theside view in FIG. 5B and is indicated by reference numeral 513.

FIGS. 5E and 5F show yet another embodiment of the Y-maze apparatus testtank of our invention. In this case, the baffle 590 is replaced by acurved perforated wall 590a. The embodiments of FIGS. 5C and 5D areindicated, overall, by reference numeral 510.

FIGS. 5C and 5D show yet another embodiment of the Y-maze apparatus ofour invention, indicated by reference numeral 520. In this case, theperforated walls 590 and 590a are replaced by perforated walls 591a and592a which contains perforated gate 593 which is removable. Fluid movespast the screen 593 and through weir 524 in exiting from the Y-maze.

FIG. 5G is a schematic diagram of the use of the Y-maze which isindicated by reference numeral 500 and which can be any of the Y-mazesas illustrated in FIGS. 5A, 5B, 5C, 5D, 5E or 5F.

The operation of the Y-maze is in two phases, phase I and phase II. Inphase I, the "preparation" and "acclimation" phase, water isrecirculated and no chemicals are tested. The test procedure is in phaseII. In phase I, fluid passes through 572a and 572b through line 572 intorecirculation bucket 571 and then through valve 574 which is open (valve577 is closed for phase I). The fluid is then pumped using pump 578through line 579 (pump 578 being also indicated as pump C). The fluidthen travels through lines 579a and 579b into sections "A" and "B" ofthe Y-maze, through lines 660a and 660b and through openings 509a and509b and lines 560a and 560b. During this procedure, valves 662 and 673are closed.

In phase II, solution is pumped from either tank 666 (the liquid beingindicated as 667) or liquid 668 is pumped from an alternative tank. Theliquids being control or test liquids. Quick connect nipples 672a and672b are interchangeable; and quick connect nipples 675a and 675b arealso interchangeable, physically. Thus, either fluid is pumped throughline 671b or is pumped through line 671a and fluid is either pumpedthrough line 663a or line 663b. In phase II, valve 577 is open and valve574 is closed.

New water 575 is pumped through line 576 past valve 577 using pump C,pump 578 through lines 579a and 579b. Then, pump B, pump 670 pumpsliquid 668 through line 669 either into line 671b or 671a depending onwhich line is connected using the quick connect nipples 672a and 672b.In any event, the fluid is pumped past valve 673 through line 660b. Bythe same token, liquid A from tank 666, the liquid being also shown byreference numeral 667 is pumped through line 665 and then either throughline 663b or line 663a depending on whether the lines are connectedusing quick connect nipple 675b or 675a. In any event, the fluid ispumped past valve 662 through line 660 past fitting 560a into section"A" of the Y-maze, the operation of which is described, supra. Theliquid 667 is pumped using pump A, pump 664. Pumps A, B and C, that ispump 664, pump 670 and pump 578 are driven by pump drives 676.

FIG. 6A sets forth a Y-maze built particularly for starfish of the priorart (Castilla, Marine Biology, Volume 12, pages 222-228 (1972)"Responses of Asterias rubens to Bivalve Prey in a Y-maze". Theapparatus indicated by reference numeral 600 is the prior art Y-mazeapparatus of the Castilla article shown on page 222 thereof.

Referring to FIG. 6B, FIG. 6B sets forth a schematic block flow diagramfor the production of Osceola Brown Sugar, produced by Osceola Farms,Inc. of Pahokee, Fla. (the headspace analysis for the Osceola BrownSugar is set forth in FIG. 27). A cane sugar tipper 401 feeds sugarcaneonto a cane feed table 402 whereupon the cane is carried on cane carrier403 into cane knives 404. The resulting product is carried on canecarrier 405 through heavy duty cane knives and through a shredder 408into magnetic tramp iron separator 407. The resulting product passesthrough juice screen 410. The unscreened material, unscreened mixedjuice, is passed through the unscreened mixed juice tank and pump 411.The screened material passes through screened juice tank and pumps 409into juice scale 449. The unscreened material is passed through pressurefed mills 412 and through imbibition juice tank and pump 415.Simultaneously, imbibition water flows out of tube 413. Simultaneously,exhaust steam from the mills and from the shredder 414 is passed throughmake up station 432 and turbonator sets 418. Boilers 422 are used toprocess bagasse coming from the bagasse conveying system 419 throughbagacillo screen 420, using the deaerater feed tank and pumps 421. Vaporbleed to vacuum pans and heaters is shown by reference numeral 423 withoverhead evaporated water 424 being condensed using condenser 425.Condensate from the tanks is collected at 426 and syrup is received atsyrup receiver and pumps 427. Syrup is pumped through the syrupclarifier 428 and sediment is collected at 429. Clarified syrup ispumped through line 430 into vacuum pan supply tanks 484.

Simultaneously, lime from source 453 is passed through lime slaker 452and screened at 451. Milk of lime tanks and pumps 450 passes theresulting product into juice liming tanks 447 where the cane sugar juiceis placed from juice heater 448. That juice is passed from juice scale449. The resulting material is placed into S.R.I. clarifier 442 equippedwith flash tank 446. The resulting clarified juice is passed throughtank and pumps 443 and then into FS clarified juice heater 434 where itis passed into the vacuum pans and heaters 423. Mud mixer and pump 440collect sediment from the clarifier 442 and the resulting pump passesmaterial into rotary vacuum filter 438 yielding filtrate from pumps 439into filtrate receivers 437. Overhead condensate is collected at 435.The condensate is passed through vacuum pump 433.

The Osceola Brown Sugar is obtained from the receiver 482 which ishooked up with magma mixer and pump 479 which receives material fromcontinuous centrifical pumps 478, which in turn receives material frommassecuite receiver and pump apparatus 476, which in turn receivesmaterial from continuous pan 475 equipped with condenser 473 andoverhead water take-off 474. Simultaneously, the seed receiver and pump485 is pumping material into continuous pan 487 equipped with condenser488 and water take-off 489. The seed receiver and pump receives materialfrom a seed pan 483 attached to vacuum pan supply tanks 484. The OsceolaBrown Sugar received at 482 is received from magma receiver 482 throughline 481 hooked up with magma mixer and pump 479. The clarified syruptank and pumps 430 passes syrup from 430 into the vacuum pan supplytanks 484. Molasses coming from line 454 is stored and that molassescoming from line 454 evolves from the molasses scale and pumps 455 whichevolves from molasses run-off tank and pump 457 attached to continuouscentrifical pumps 456 working with reheater 459 and meter and pump 458.The reheater 459 is worked continuously with the continuous verticalcrystalizers 460 which is hooked up with massecuite receiver and pump466 associated with continuous pan 463 having attached thereto condenser461 and water takeoff 462. Continuous pan 463 is attached to "C" seedreceiver and pump 467 receiving material from seed pan 465 which ishooked up with condensers 468 and water take-off 469. Vacuum supplytanks 464 are hooked up with both the "C" seed pan 465 and thecontinuous pan 463. The seed pan 465 is also hooked up with condensers468 and water take-off 469 as is seed pan 471. Seed pan 471 is hooked upwith seed receiver and pump 472. Seed pan 471 is also hooked up withvacuum pan supply tanks 470 which in turn is hooked up to molassesrun-off tank and pump 477 hooked up in turn with continuous centrificalpumps 478 and magma mixer and pump 479.

Multiple effect evaporator 431 causes the creation of syrup received at427 which is then pumped into syrup clarifier 428.

The material received from the "B" magma receiver 482 is crystalized andthe Osceola Brown Sugar is in fact the crystals received from 482.

FIG. 27 (also referred to, infra) is a headspace analysis of substancestrapped on TENAX® and is a gas chromatograph for the headspace for theOsceola Brown Sugar produced by Osceola Farms Inc. of Pahokee, Fla.according to the process set forth in the detailed description of FIG.6B, supra.

FIG. 7A is a perspective diagrammatic view of the apparatus of FIG. 2A.FIG. 7C is a perspective view in diagrammatic form of the apparatus ofFIG. 3 incorporating FIGS. 3A, 3B and 3C.

With respect to each of the figures showing the video camera orcamcorder described, supra, a useful video camera is a SONY® VideoCamera Recorder Hi8, Model CCD-TR101 (Video Hi8 "Handycam").

FIG. 8 sets forth a graph showing the response "R" on the "Y" axisversus:

     -log.sub.10 C!

(with C being in gram moles per liter) for N-acetyl-D-Glucosamine, anepimeric mixture of compounds having the structures: ##STR91##

The graph indicated by reference numeral 800 is a linear regressiongraph drawn through each of the four points which are data points 801,802, 803 and 804. The graph indicated by reference numeral 800a is aparabola drawn through each of the four points which are data points801, 802, 803 and 804.

By the same token, in FIG. 9 the graph of:

     -log.sub.10 C!

versus "R" for the materials TALIN® (shown by linear regression graph901 using data points 959, 960 and 961 and shown by the parabola 901a);S-methyl methionine sulfonium chloride having the structure: ##STR92##(using linear regression graph 902 and parabola 902a and data points962, 963 and 966); and D-Glucosamine, an epimeric mixture of thecompounds having the structures: ##STR93## (using linear regressiongraph 900 and data points 950 and 951) for the mean of the data pointsand graph 900a and data points 952 and 953 for the median of the datapoints. The equation defining the line indicated by reference numeral900a is:

    R=-1/3 log.sub.10 C!=log.sub.10 C.sup.-1/3.

The equation defining the line indicated by reference numeral 900 is:##EQU25##

By the same token, in FIG. 10 the graph of:

     -log.sub.10 C!

versus response "R" is for two materials, propiothetin (bromide) havingthe structure: ##STR94## (shown by linear regression graph 1002 andparabola 1002a and data points 1060, 1061 and 1062) and trimethyl amineoxide hydrate having the structure: ##STR95## (shown by linearregression graph 1000 and data points 1050 and 1051).

By the same token, FIG. 11 shows a graph of:

     -log.sub.10 C!

versus response "R" for use in the static test tank of FIG. 2A for thespecies Penaeus vannamei for the racemic mixture of 1-octen-3-ol havingthe structure: ##STR96## an isomeric mixture of the compounds having thestructures: ##STR97## (shown by linear regression graph 1100 andparabola 1100a and data points 1150, 1152 and 1151) and for guanidinehaving the structure: ##STR98## (shown by linear regression graph 1101and data points 1160 and 1161).

By the same token, FIG. 12 is a graph of:

     -log.sub.10 C!

(with C in grams per liter) versus "R" for TALIN®. The graph indicatedby reference numeral 1200 is a graph for a straight line of:

     -log.sub.10 C!

versus "R" directly through data points 1250 and 1251.

By the same token, FIG. 13, the graph of:

     -log.sub.10 C!

versus "R" is for two materials:dimethyl sulfoxide having the structure:##STR99## (indicated by the linear regression graph 1304 and theparabola 1304a and the data points 1305, 1306 and 1307) and formethional having the structure: ##STR100## (indicated by the linearregression graph 1300 and the parabola 1300a and data points 1301, 1302and 1303). A mathematical model for the parabola 1300a is:

    R=-0.0533(log.sub.10 C).sup.2 -1.23 log.sub.10 C-2.25

which is of the form:

    R=α(log.sub.10 C).sup.2 +βlog.sub.10 C+γ.

A mathematical model for the parabola 1304a is:

    R=0.074(log.sub.10 C).sup.2 +1.83(log.sub.10 C)+14.47

which is of the form:

    R=α(log.sub.10 C).sup.2 +βlog.sub.10 C+γ.

By the same token, the graph of FIG. 14 of:

     -log.sub.10 C!

versus "R" is for the material, a 50:50 mole:mole mixture of skatolehaving the structure: ##STR101## and indole having the structure:##STR102## (with linear regression graph 1400 and parabola 1400a throughdata points 1401, 1402 and 1403).

The graph of FIG. 15 is also for:

     -log.sub.10 C!

versus "R" (response) for three materials for the species Penaeussetiferus tested in a static holding tank. The graph indicated byreference numeral 1500 is a regression parabolic curve involving datapoints 1501, 1502, 1503 and 1504 and is for aqueous ammonia. The graphindicated by reference numeral 1510 is a linear regression graph foracetic acid and its data points are indicated by reference numerals1511, 1512 and 1513. The graph indicated by reference numeral 1500a is aparabolic curve for acetic acid directly through the data points 1511,1512 and 1513.

The graph indicated by reference numeral 1520 is a linear regressiongraph for aqueous ammonium chloride and its data points are indicated byreference numerals 1521, 1522 and 1523. The graph indicated by referencenumeral 1520a is a parabola directly through the data points indicatedby reference numerals 1521, 1522 and 1523.

By the same token, FIG. 16 is a graph of:

     -log.sub.10 C!

versus response "R" using a static holding tank testing apparatus andinvolving the species Penaeus setiferus for the substances betainehaving the structure: ##STR103## (indicated by the linear regressiongraph 1610 and the data points 1611, 1612 and 1613); glycine (indicatedby the linear regression graph 1600 and the data points 1601, 1602 and1603); and for aspartate ion (sodium aspartate in solution; indicated bylinear regression graph 1620 and the data points 1621, 1622 and 1623).

By the same token, FIG. 17 is a graph of:

     -log.sub.10 C!

versus response "R" using the static holding tank apparatus of FIG. 2involving the species Penaeus setiferus. The graph indicated byreference numeral 1710 is for aqueous glucose and is a straight linethrough data points 1711 and 1712. Graph 1700 is a linear regressiongraph for taurine having the structure: ##STR104## using data points1701, 1702 and 1703.

A chemoreception static test tank embodiment is set forth in theillustration in FIG. 18, in perspective. Various locations at the bottomof test tank 1800 (with the bottom indicated by reference numeral 1801)are set forth in the test tank for reference purposes. Point "a" isindicated by reference numeral 1803. Point "b" is indicated by referencenumeral 1804. Point "c" is indicated by reference numeral 1805. Point"d" is indicated by reference numeral 1806. Point "E" is indicated byreference numeral 1807. Point "F" is indicated by reference numeral1808. Point "G" is indicated by reference numeral 1809. Point "H" isindicated by reference numeral 1810. The chemical injection point in thetank is indicated by reference numeral 1802. The member of the Penaeusgenus of the Class Crustacea will move from location "b" towards any ofthe other locations over a given period of time with a given feedinserted at 1802. That movement is recorded and is a function of thegrade "G" or "response", "R", for the particular stimulant at theparticular concentration involved at injection point 1802.

Using the flow-through equipment of FIGS. 3 and 3B, a graph of flow rate(in ml per minute) versus response for the material: "glucose+naturalextract+salt water" (described, supra) is indicated by linear regressiongraph 1900 using data points 1901, 1902 and 1903.

FIG. 20 sets forth three graphs for aqueous ammonium acetate of:

     -log.sub.10 C!

versus response "R" for three different flow rates using theflow-through apparatus of FIGS. 3 and 3B. The graph indicated byreference numeral 2000 is for a flow rate of 125 ml/minute and is astraight line through data points 2001 and 2002. The graph for a flowrate of 100 ml/minute is indicated by reference numeral 2020 and is astraight line through data points 2021 and 2022. The graph indicated byreference numeral 2010 is for a flow rate of 75 ml/minute and is astraight line through data points 2011 and 2012.

When using as test materials the ammonium chloride, ammonium acetate,aqueous ammonia and acetic acid as set forth, supra, a number ofequilibria exist: ##STR105##

For the equation:

    NH.sub.4.sup.+ +H.sub.2 O⃡H.sup.+ +NH.sub.4 OH,

the relationship: ##EQU26## exists. For the equilibrium:

    NH.sub.4 OH⃡NH.sub.4.sup.+ +OH.sup.-

the relationship:

    K.sub.i =1.8×10.sup.-5

exists.

For the equilibrium:

    H.sub.2 O⃡H.sup.+ +OH.sup.-

the relationship:

    K.sub.w =10.sup.-14

exists.

For the equilibrium: ##STR106## the relationship:

    K.sub.5 =1.8×10.sup.-5

exists.

When ammonium acetate having the structure: ##STR107## hydrolyzes,according to the equilibrium : ##STR108## the acetic acid and aqueousammonia further hydrolyze to form acetate ions and ammonium ions and therelationship for the hydrolysis constant:

    K.sub.hAA

for ammonium acetate is: ##EQU27##

For example, at a concentration of 10⁻⁹ for the ion:

     NH.sub.4.sup.+ !

the amount of hydrolysis to aqueous ammonia is negligible since thehydrolysis constant is 5.5×10⁻¹⁰.

At an initial concentration,

     NH .sub.4 OH!

of 10⁻¹², the amount of ionization to ammonium ion is negligible sinceK_(i) is 1.8×10⁻⁵.

Since all of the above equilibria are in saline solution containingchloride cations and metal anions such as sodium ion, many ion pairsexist and the standard equilibria and mathematical relationships areaccordingly altered.

Referring to the graph of FIG. 21, for:

     -log.sub.10 C!

versus response for N-acetyl-D-Glucosamine, an epimeric mixture ofcompounds having the structures: ##STR109## the linear regression graph,a straight line, is indicated by reference numeral 2110 and the datapoints for said regression graph are indicated by reference numerals2112, 2114 and 2116. The flow-through tank used is that set forth inFIGS. 3 and 3B described, supra, and the species tested is Penaeusvannamei.

FIG. 22, showing the relationship of:

     -log.sub.10 C!

versus response "R", shows two different graphs using three data points2212, 2214 and 2216 for methional having the structure: ##STR110## Theregression graph indicated by reference numeral 2210 is in the shape ofa parabola and the mathematical model therefor is as follows:

    R=0.167(log.sub.10 C).sup.2 +3.07 (log.sub.10 C)+26.092.

The linear regression graph indicated by reference numeral 2211 is for astraight line. The apparatus used is the flow-through apparatus of FIGS.3 and 3B and the species involved is Penaeus vannamei.

Table VI below sets forth mean response versus number of species ofPenaeus vannamei in the group versus the particular chemical involvedand its concentration. Table VI is as follows:

                  TABLE VI                                                        ______________________________________                                                         Number of                                                                              Average Response Using                                               Species  the Mathematical Model:                             Chemical and Concentration                                                                     in Group G.sub.2 = 100 - 10θ                           ______________________________________                                        " " (Solution of FRITZ ®                                                                   16       5.82                                                salt in water, 35 ppt)                                                        Natural extract: 8        17.33                                               Glucose at 10.sup.-3 M                                                        N-acetyl-D-Glucosamine at                                                                      8        17.85                                               a concentration of 10.sup.-12 M                                               in 35 ppt aqueous FRITZ ®                                                 salt solution                                                                 N-acetyl-D-Glucosamine at                                                                      8        22.44                                               a concentration of 10.sup.15 M                                                in 35 ppt aqueous FRITZ ®                                                 salt solution                                                                 N-acetyl-D-Glucosamine at                                                                      8        25.41                                               a concentration of 10.sup.-18 M                                               in 35 ppt aqueous FRITZ ®                                                 salt solution                                                                 Propiothetin (bromide) at                                                                      7        2.89                                                10.sup.9 M in aqueous FRITZ ®                                             salt solution at 35 ppt                                                       S-Methyl methionine                                                                            8        7.69                                                sulfonium chloride at 10.sup.9 M                                              (in aqueous FRITZ ® salt                                                  solution at 35 ppt)                                                           TALIN ® at 10.sup.-5 mg/L (of                                                              8        27.28                                               aqueous FRITZ ® salt                                                      solution at 35 ppt)                                                           Trimethyl amine oxide                                                                          7        19.72                                               hydrate at 10.sup.9 M (in                                                     aqueous FRITZ ® salt                                                      solution at 35 ppt)                                                           TASTONE ® 900 (Bakers Yeast                                                                8        20.20                                               Extract, spray-dried                                                          manufactured by the Red                                                       Star Specialty Products                                                       Inc. of 433 East Michigan                                                     Street, Milwaukee,                                                            Wisconsin 53202) (10 mg/L                                                     of FRITZ ® salt solution at                                               35 ppt)                                                                       Methional at 10.sup.9 M (in                                                                    8        18.43                                               aqueous FRITZ ® salt                                                      solution at 5 ppt)                                                            Methional at 10.sup.-12 M (in                                                                  8        13.29                                               aqueous FRITZ ® salt                                                      solution at 5 ppt)                                                            Methional at 10.sup.15 M (in                                                                   7        24.94                                               aqueous FRITZ ® salt                                                      solution at 5 ppt)                                                            Dimethyl sulfoxide at                                                                          8        43.40                                               10.sup.-12 M (in aqueous FRITZ ®                                          salt solution at 5 ppt)                                                       ______________________________________                                         Note: The aqueous FRITZ ® salt, described in detail, supra, is at a       level in water of 35 parts per thousand in each of the compositions set       forth in Table VI.                                                       

FIG. 23 is a liquid chromatogram profile for TALIN® (trademark of Tateand Lyle Limited of the United Kingdom), a mixture of Thaumatin I,Thaumatin II and Thaumatin B. (Conditions: S-Sepharose column operatingat 7 ml per minute; gradient: 0-25 mM NaCl (2×750 ml) fraction size: 45ml.) The peak indicated by reference numeral 2316 is for that part ofTALIN® which is known as "Thaumatin I" as described in U.S. Pat. No.5,221,624 issued on Jun. 22, 1993, the specification for which isincorporated by reference herein. The Thaumatin I can also be shownusing the symbolism:

     Lys.sup.46, Asp.sup.113, Asp.sup.137 !

where "Lys" stands for "lysine" moiety; and "Asp" stands for an"Aspartic acid" moiety. The peaks indicated by reference numerals 2312and 2314 are for "Thaumatin B" and "Thaumatin II" as described in U.S.Pat. No. 4,771,000, the specification for which is incorporated byreference herein.

FIG. 24 is the high pressure liquid chromatography profile for the sameTALIN® as set forth concerning the description of FIG. 23.

FIG. 25 is a graph of:

     -log.sub.10 C!

versus response "R" (using the static tank test apparatus of FIGS. 2Aand 7A) for 2-methyl-3-(methyldithio)furan having the structure:##STR111## The graph indicated by reference numeral 2300 and byreference numeral 2300a are "regression" graphs for a "cubic" equation.The cubic equation is of the form:

    R=α(-log.sub.10 C).sup.3 +β(-log.sub.1o C).sup.2 +γ(-log.sub.10 C)+δ

and is in fact:

    R=0.0096(log.sub.10 C).sup.3 -0.74 (log.sub.10 C).sup.2 -172.6 log.sub.10 C-2604.63

wherein α, β, γ and δ are constants.

FIG. 26 is a graph of:

     -log.sub.10 C!

versus response "R" using the static tank test apparatus of FIGS. 2A and7A for "Osceola Brown Sugar", a product which is raw sugar containingimpurities produced by Osceola Farms Inc. of Pahokee, Fla. The graphindicated by reference numeral 2400 is a linear regression graph, astraight line. The graph indicated by reference numeral 2400' is for ahyperbola and has the mathematical model: ##EQU28## which is of theform: ##EQU29## wherein α, β and γ are constants. The graph indicated byreference numeral 2400' is directly through the data points indicated byreference numerals 2401', 2402' and 2403'.

A duplicate set of points, 2401", 2402" and 2403" for the Osceola BrownSugar yields an exponential equation for the exponential graph indicatedby reference numeral 2400", having the mathematical model:

    R=3.4+5.33e.sup.+2.4 log.sbsp.10.sup.C.

FIG. 27 is a headspace analysis trapped on TENAX® and is a gaschromatograph for the headspace for the Osceola Brown Sugar produced byOsceola Farms Inc. of Pahokee, Fla.

The peak indicated by reference numeral 2705 is for carbon dioxide. Thepeak indicated by reference numeral 2706 is for dimethyl sulfide havingthe structure: ##STR112## The peak indicated by reference numeral 2707is for acetic acid having the structure: ##STR113## The peak indicatedby reference numeral 2702 is for dimethyl sulfoxide having thestructure: ##STR114## The peak indicated by reference numeral 2701 isfor a mixture of 2,3-dimethyl pyrazine and 2,5-dimethyl pyrazine havingthe structures, respectively: ##STR115## The peak indicated by referencenumeral 2703 is for 2,3,5-trimethyl pyrazine having the structure:##STR116## The peak indicated by reference numeral 2704 is for paravinylguiacol.

FIG. 28 is a "VENN" diagram showing the sets of incitants, stimulants,attractants and excitants for members of the Penaeus genus of the ClassCrustacea. The overlapping area 2801 is that where particular substancesare simultaneously incitants, stimulants, attractants and excitants formembers of the Penaeus genus of the Class Crustacea. The symbol "I" isfor incitants. The symbol "S" is for stimulants. The symbol "A" is forattractants. The symbol "E" is for excitants.

FIG. 29 is a graph of:

     -log.sub.10 C!

versus "response" using flow-through apparatus of FIGS. 3 and 3B forN-acetyl-D-Glucosamine, an epimeric mixture of compounds having thestructures: ##STR117## The graph indicated by reference numeral 2900 isfor the mean of data points and these mean data points are indicated byreference numerals 2901, 2902 and 2903. The mathematical model for thegraph indicated by reference numeral 2900 is as a "hyperbola", to wit:##EQU30## or an exponential equation, to wit: ##EQU31## The graphindicated by reference numeral 2910 is for a straight line for themedian data points and these median data points are represented bypoints 2911, 2912 and 2913. The mathematical model for the graphindicated by reference numeral 2910 is:

    R=-41.98-5.68 log.sub.10 C.

FIG. 30 sets forth two graphs for the relationship of:

     -log.sub.10 C!

versus "R" for methional having the structure: ##STR118## using theflow-through apparatus of FIGS. 3 and 3B. The graph indicated byreference numeral 3000 is a straight line graph for "mean" data points3001, 3002 and 3003 and has the mathematical model:

    R= -6.71-2.44 log.sub.10 C!.

The graph indicated by reference numeral 3010 is for the "median" datapoints 3011, 3012 and 3013 and is an exponential equation, to wit:##EQU32##

FIG. 31 is a pair of graphs, a "median" graph and a "mean" graph for therelationship of:

     -log.sub.10 C!

versus "R" using the flow-through tank apparatus of FIGS. 3 and 3B for a50:50 mole:mole mixture of skatole having the structure: ##STR119## andindole having the structure: ##STR120## The "median" graph is indicatedby reference numeral 3110 for the "median" data points 3111, 3112 and3113 and is a parabola having the equation:

    R=-2.34 log.sub.10 C!.sup.2 -58.77 log.sub.10 C-338.

The "mean" data points are indicated by the parabola 3100 through "mean"data points 3101, 3102 and 3103. The "mean" data point parabola has theequation:

    R=-0.56 log.sub.10 C!.sup.2 -14.33 log.sub.10 C-62.

FIG. 32 sets forth a pair of graphs, a "mean" data point graph and a"median" data point graph of:

     -log.sub.10 C!

versus "R" in the flow-through tank apparatus of FIGS. 3 and 3B formethionine having the structure: ##STR121## The graph indicated byreference numeral 3210 is for the median data points through points 3211and 3212. The graph indicated by reference numeral 3200 is for the"mean" data points through data points 3201 and 3202.

FIG. 33 sets forth a pair of graphs, a "mean" data point graph and a"median" data point graph for the relationship of:

     -log.sub.10 C!

versus response "R" using static tank apparatus of FIGS. 2A and 7A foryeast extract. The graph indicated by reference numeral 3310 is for the"median" data points and goes directly through points 3311 and 3312. Thegraph indicated by reference numeral 3300 is for the "mean" data pointsand goes directly through points 3301 and 3302.

FIG. 34 sets forth two "response as a function of concentration" graphssetting forth data for the response "R" on the "X" axis versus:

     -log.sub.10 C!

on the "Y" axis (and "C" being concentration in gram moles per liter)for N-acetyl-D-Glucosamine, an epimeric mixture of compounds having thestructures: ##STR122## in the static tank testing apparatus of FIG. 7Bas against the Procambrus clarkii (crawfish) species of the ClassCrustacea. The graph indicated by reference numeral 601' (a "gamma"function) is for the means of the responses "R" versus:

     -log.sub.10 C!.

The graph indicated by reference numeral 651' (a regression curve) isfor the medians of responses "R" versus:

     -log.sub.10 C!.

The graph indicated by reference numeral 601' is a graph described bythe mathematical model:

    R=5.0014- log.sub.10 C!.sup.2 (0.0084+0.5169e.sup.+0.45 log.sbsp.10.sup.C!),

drawn through each of the points which are data points 606', 602', 603',604' and 605'. The graph indicated by reference numeral 651' is aregression curve described by the mathematical model:

    R=0.7874 -log.sub.10 C!.sup.-3/2 -0.0001 log.sub.10 C!.sup.2 +3.041

and is for the median points, 656', 657', 655', 654', 653' and 652'.

With respect to the "mean" line, if point 607' were taken intoconsideration then a second mean line ("mean line" II) could be drawndescribed according to the mathematical model:

    R=4.6574+0.0018 log.sub.10 C!.sup.3 +0.0322 log.sub.10 C!.sup.2 +0.0907 log.sub.10 C!,

a cubic equation. The mean line II is indicated by reference numeral601".

FIG. 35 sets forth two "response as a function of concentration" graphs("gamma" function and hyperbolic function) of:

     -log.sub.10 C!

versus response "R" with:

     -log.sub.10 C!

being on the "Y" (and "C" being concentration in gram moles per liter)and "R" being on the "X" axis for the testing of the substance trimethylamine oxide hydrate having the structure: ##STR123## in the static tanktesting apparatus of FIG. 7B against the Procambrus clarkii species ofthe Class Crustacea. The graph indicated by reference numeral 701' (a"gamma" function) is for the means of responses "R" versus:

     -log.sub.10 C!

and is defined by the mathematical model:

    R=(-6×10.sup.-4) log.sub.10 C!e.sup.-0.2569 log.sbsp.10.sup.C! +0.0207 log.sub.10 C!+3.5607.

The graph indicated by reference numeral 751' (a hyperbolic function) isfor the medians of responses "R" versus:

     -log.sub.10 C!

and is defined by the mathematical equation:

    R=-2.6824 sin h -0.1{log.sub.10 C}!-0.3680 log.sub.10 C!+3.1913.

The data points for the graph of the means 701' are indicated byreference numerals 702', 703', 704' and 705'. The data points for thegraph of the medians which graph is indicated by reference numeral 751',are indicated by reference numerals 752', 753', 754' and 755'.

FIG. 36 sets forth two "response as a function of concentration" graphs(the means of response graph being indicated by reference numeral 801',a "gamma" function and the medians of response graph being indicated byreference numeral 851' (also a "gamma" function)) of:

     -log.sub.10 C!

on the "Y" axis versus response "R" on the "X" axis with "C" being ingram moles per liter, for the substance, Osceola Brown Sugarmanufactured by Osceola Farms Inc. of Pahokee, Fla. (analysis ofheadspace as trapped on TENAX® is set forth in FIG. 27) in the statictank testing apparatus of FIG. 7B as against the Procambrus clarkiispecies of the Class Crustacea.

The means of response graph indicated by reference numeral 801' isdefined according to equation:

    R=(-0.0082) log.sub.10 C!.sup.2 e.sup.-0.0573 log.sbsp.10.sup.C! -0.0066 log.sub.10 C!.sup.2 +6.3210

and is for the data points indicated by reference numerals 802', 803',804' and 805'.

The medians of response graph 851 is defined according to themathematical model:

    R=(-1.5542×10.sup.4){log.sub.10 C}.sup.2 e.sup.+2.1941 log.sbsp.10.sup.C! -0.1613 log.sub.10 C!.sup.2 +11.2387

and is for the data points indicated by reference numerals 852, 853, 854and 855.

FIG. 37 sets forth two "response as a function of concentration" graphssetting forth data for the response "R" on the "X" axis versus:

     -log.sub.10 C!

on the "Y" axis (and "C" being concentration in gram moles

per liter) for "METHYLOXYCYCLOSULFIDE™" (a trademark of InternationalFlavors & Fragrances Inc.) also called "MOS", the compound having thestructure: ##STR124## in the static tank testing apparatus of FIG. 7B asagainst the Procambrus clarkii (crawfish) species of the ClassCrustacea. The graph indicated by reference numeral 3700 (a "gamma"function) is for the median of the response "R" versus:

     -log.sub.10 C!.

The graph indicated by reference numeral 3710 (a "gamma" function) isfor the means of the response "R" versus:

     -log.sub.10 C!.

The graph indicated by reference numeral 3700 is a graph described bythe mathematical model:

    R=152.9674-0.732 log.sub.10 C!.sup.2 e.sup.0.0485 log.sbsp.10.sup.C +12.433 log.sub.10 C!

drawn through each of the points which are data points 3701, 3702, 3703and 3704. The graph indicated by reference numeral 3710 is a graphdescribed by the mathematical model:

    R=0.1302 log.sub.10 C!.sup.2 e.sup.0.092 log.sbsp.10 C!-0.3697 log.sub.10 C!-5.0549

drawn through each of the points which are data points 3711, 3712, 3713and 3714.

FIG. 38A is a series of bar graphs showing percent weight gain during asix-week feed trial for members of the Penaeus genus of the ClassCrustacea using Diets A, P, J and C (described, supra). The bar graphindicated by reference numeral 3801A is for the use of Diet A. The bargraph indicated by reference numeral 3802C is for Diet C. The bar graphindicated by reference numeral 3803J is for Diet J. The bar graphindicated by reference numeral 3804P is for Diet P. Percent weight gainfor the members of the Penaeus genus of the Class Crustacea is set forthon the "Y" axis. The particular diet is set forth on the "X" axis.

FIG. 38B is a series of bar graphs showing average growth in six weeks(in grams) for members of the Penaeus genus of the Class Crustacea usingeach of Diets A, C, J and P, each of which is described, supra; Diet Cbeing the "control". The average growth for the six-week period is setforth in grams on the "Y" axis. The bar graph indicated by referencenumeral 3810A is for Diet A. The bar graph indicated by referencenumeral 3811C is for Diet C. The bar graph indicated by referencenumeral 3812J is for Diet J. The bar graph indicated by referencenumeral 3813P is for Diet P.

FIG. 38C is a series of bar graphs setting forth the total number of"hits" over a six-week period by members of the Penaeus genus of theClass Crustacea. The total number of "hits" is set forth on the "Y"axis. The specific diet used is set forth on the "X" axis. The bar graphindicated by reference numeral 3820A is for Diet A. The bar graphindicated by reference numeral 3821C is for Diet C. The bar graphindicated by reference numeral 3822J is for Diet J. The bar graphindicated by reference numeral 3823P is for Diet P.

FIG. 39 relates to the data summarized in FIGS. 38A, 38B and 38C. FIG.39 sets forth in a "exact fit" curve and a linear regression lineplotting the number of "hits" (in units of hundreds of "hits") versusthe weight gain for members of the Penaeus genus of the Class Crustacea.The point indicated by reference numeral 3904J is for Diet J. The pointindicated by reference numeral 3903P is for Diet P. The point indicatedby reference numeral 3902C is for Diet C. The point indicated byreference numeral 3901A is for Diet A. The "exact fit" curve isindicated by reference numeral 3900. The linear regression line isindicated by reference numeral 3910.

The "exact fit" curve indicated by reference numeral 3900 is definedaccording to the mathematical model: ##EQU33## The linear regressionline 3910 is defined according to the mathematical model:

    H=66.67(ΔW)-44.33.

wherein weight gain is shown by the symbol: ΔW and wherein the number of"hits" is indicated by: H .

FIG. 40 is a graph showing the differences between the control (Diet C)and the other diets (Diets J, A and P) the growth data for which issummarized in FIGS. 38A, 38B and 38C and set forth, numerically, supra.

FIG. 40 is a graph showing on the "Y" axis the difference in the numberof "hits" between the control diet and Diets J, A and P versus thechange in the amount of growth between the control, Diet C, and Diets A,J and P indicated by the symbol: Δ(ΔW). The graph indicated by referencenumeral 4000 is a linear regression line through points 4001, 4002 and4003. The data point indicated by reference numeral 4003 sets forth thedifferences between the control ("zero") and Diet A. The point indicatedby reference numeral 4002 is for the difference between the control andDiet P. The point indicated by reference numeral 4001 is for thedifference between the control and Diet J. Linear regression line 4000is defined according to the mathematical model:

    ΔH=5333.33(Δ ΔW!)-180.

FIG. 41 is a series of graphs showing the differences between Diets A, Jand P (described, supra) and Diet C, the control, plotting change inweight gain on the "X" axis versus percent "active" on the "Y" axis withthe "active" being either the mixture, to wit:

N-acetyl-D-Glucosamine;

dimethyl sulfoxide; and

methional

hereinafter referred to as "TRIO" or Diet J which is the compound havingthe structure: ##STR125## (hereinafter referred to as "MOS").

The graph indicated by the reference numeral 4100 is for Penaeusvannamei. The data point indicated by reference numeral 4101 is for DietJ and the data point indicated by reference numeral 4102 is for Diet Acontaining 37.5% "TRIO".

The line indicated by reference numeral 4110 is for Penaeus aztecususing "TRIO". The data point indicated by reference numeral 4111 is forDiet P containing all "TRIO". The data point indicated by referencenumeral 4112 is for Diet A which contains 37.5% "TRIO".

The line indicated by reference numeral 4120 is for Penaeus vannameiusing Diet J, "METHYLOXYCYCLOSULFIDE™". The point indicated by referencenumeral 4121 is for Diet J containing 100% "MOS". The point indicated byreference numeral 4122 is for the amount of "MOS" contained in Diet Awhich is 12.5% (there being seven other components).

The line indicated by reference numeral 4130 is for Penaeus aztecususing the Diet J "MOS". The point indicated by reference numeral 4131 isfor Diet J, which is all "MOS". The point indicated by reference numeral4132 is for the "MOS" in Diet A, the "MOS" being 12.5% of Diet A. Diet Acontains seven other components in equal amounts.

The graph indicated by reference numeral 4100 is defined according tothe mathematical model:

    A=(431.03)Δ(ΔW)+132.33.

or the mathematical model: ##EQU34##

The graph indicated by reference numeral 4110 is defined according tothe mathematical model: ##EQU35##

The graph indicated by reference numeral 4120 is defined according tothe mathematical model: ##EQU36##

The graph indicated by reference numeral 4130 is defined according tothe mathematical model: ##EQU37##

The data set forth in FIGS. 38A, 38B and 38C is also described in FIGS.42 and 43.

Accordingly, FIG. 42 is a three-dimensional graph showing on the "Y"axis the change in the change of weight gain indicated by the symbol:Δ(ΔW); on the "X" axis log₁₀ (of the change in the number of "hits")indicated by the symbol: log₁₀ (ΔH); and on the "Z" axis by log₁₀ Aindicated by the symbol: log₁₀ A; wherein "A" is the percent "active".The same coordinates are used for FIG. 43. FIG. 42 has as the "active"material, "TRIO". The graph indicated by reference numeral 4200 is drawnthrough data points 4201 and 4202. Data point 4201 indicates Diet Pcontaining 100% "TRIO". The data point indicated by reference numeral4202 indicates "TRIO" in Diet A; that is, 37.5% "TRIO" which Diet Acontains.

In FIG. 43, the line indicated by reference numeral 4300 is drawnthrough data points 4301 and 4302 and is for Diet J. Reference numeral4301 is for 100% Diet J containing the compound having the structure:##STR126## Reference numeral 4302 is for "MOS" contained in Diet A, inan amount of 12.5%.

The graph indicated by reference numeral 4200 in FIG. 42 is definedaccording to the mathematical model:

    -1.7013 log.sub.10 (ΔH)!+1.1254 Δ(ΔW)!+3.0019 log.sub.10 A!=1

or the mathematical model: ##EQU38##

The graph indicated by reference numeral 4300 in FIG. 43 is definedaccording to the mathematical model:

    3.647 log.sub.10 (ΔH)!-1.636( Δ(ΔW)!)+0.345 log.sub.10 A!=10.

FIG. 44 sets forth two "response as a function of concentration" graphssetting forth data for the response "R" on the "X" axis versus:

     -log.sub.10 C!

on the "Y" axis (and "C" being concentration in gram moles per liter)for "OXYCYCLOTHIONE-030™", (trademark of International Flavors &Fragrances Inc.) having the structure: ##STR127## in the static tanktesting apparatus of FIG. 2A as against the Penaeus vannamei species ofthe Penaeus genus of the Class Crustacea. The graph indicated byreference numeral 4400 is for the medians of the responses "R" versus:

     -log.sub.10 C!.

The graph indicated by reference numeral 4410 is for the means ofresponses "R" versus:

     -log.sub.10 C!.

The mean line 4410 is drawn through data points 4411 and 4412. Themedian line 4400 is drawn through data points 4401 and 4402.

The mean line 4410 is defined according to the mathematical model:

    R=7.3792+0.4167 log.sub.10 C!.

The median line indicated by reference numeral 4400 is defined accordingto the mathematical model:

    R=8.5+0.5 log.sub.10 C!.

What is claimed is:
 1. An enhanced, liquid feed composition for a memberof the Class Crustacea comprising in intimate admixture:(a) water; (b)sodium chloride; (c) a basic fish meal-containing feed composition for amember of the Class Crustacea, and intimately admixed therewith; (d) anattracting and/or stimulating and/or inciting and/or excitingconcentration and quantity of at least one feed enhancing substance in aconcentration of from about 10⁻⁴ gram moles per liter down to about10⁻²¹ gram moles per liter selected from the group consisting of:(i)N-acetyl-D-Glucosamine, an epimeric mixture of compounds having thestructures: ##STR128## (ii) S-methyl methionine sulfonium halidesdefined according to the structure: ##STR129## wherein X is a chlorideor bromide anion; (iii) methionine having the structure: ##STR130## (iv)trimethyl amine oxide hydrate having the structure: ##STR131## (v)1-octen-3-ol having the structure: ##STR132## (a mixture of isomershaving the structures: ##STR133## (vi) methional having the structure:##STR134## (vii) dimethyl sulfoxide having the structure: ##STR135##(viii) a 50:50 mole:mole mixture of skatole/indole, skatole having thestructure: ##STR136## and indole having the structure: ##STR137## (ix)propionthetin halides having the structure: ##STR138## wherein X is achloride or bromide anion; (x) ammonium chloride; (xi) ammonium acetate;(xii) acetic acid; (xiii) glucose; (xiv) "raw sugar", a mixture ofsucrose and impurities; (xv) Thaumatin; (xvi)2-methyl-3-(methyldithio)furan having the structure: ##STR139## (xvii)aqueous ammonia; (xviii) yeast extract; (xix) D-Glucosamine, an epimericmixture of compounds having the structures: ##STR140## and (xx)2-methyl-3-furanthiol having the structure: ##STR141##
 2. The enhancedfeed composition of claim 1 wherein the feed enhancing substance isN-acetyl-D-Glucosamine, an epimeric mixture having the structures:##STR142##
 3. The enhanced feed composition of claim 1 wherein the feedenhancing substance is at least one S-methyl methionine sulfonium halidehaving the structure: ##STR143## wherein X is a chloride anion or abromide anion.
 4. The enhanced feed composition of claim 1 wherein thefeed enhancing substance is methionine having the structure: ##STR144##5. The enhanced feed composition of claim 1 wherein the feed enhancingsubstance is trimethyl amine oxide hydrate having the structure:##STR145##
 6. The enhanced feed composition of claim 1 wherein the feedenhancing substance is racemic 1-octen-3-ol having the structure:##STR146## a mixture of isomers having the structures: ##STR147##
 7. Theenhanced feed composition of claim 1 wherein the feed enhancingsubstance is methional having the structure: ##STR148##
 8. The enhancedfeed composition of claim 1 wherein the enhancing feed substance isdimethyl sulfoxide having the structure: ##STR149##
 9. The enhanced feedcomposition of claim 1 wherein the enhancing feed substance is a 50:50mole:mole mixture of skatole/indole, skatole having the structure:##STR150## and indole having the structure: ##STR151##
 10. The enhancedfeed composition of claim 1 wherein the feed enhancing substance is atleast one propionthetin halides defined according to the structure:##STR152## wherein X is a chloride or bromide anion.
 11. The enhancedfeed composition of claim 1 wherein the feed enhancing substance is rawsugar, a mixture of sucrose and impurities.
 12. The enhanced feedcomposition of claim 1 wherein the feed enhancing substance isThaumatin.
 13. The enhanced feed composition of claim 1 wherein the feedenhancing substance is 2-methyl-3-(methyldithio)furan having thestructure: ##STR153##
 14. The composition of claim 1 wherein the feedenhancing substance is a mixture of:(i) N-acetyl-D-Glucosamine, anepimeric mixture of compounds having the structures: ##STR154## (ii)dimethyl sulfoxide having the structure: ##STR155## and (iii) methionalhaving the structure: ##STR156##
 15. The composition of claim 1 whereinthe feed enhancing substance is a mixture of:(i)2-methyl-3-(methyldithio)furan having the structure: ##STR157## (ii)N-acetyl-D-Glucosamine, an epimeric mixture of compounds having thestructures: ##STR158## (iii) dimethyl sulfoxide having the structure:##STR159## (iv) brown sugar; (v) a 50:50 mixture of skatole and indole,with the skatole having the structure: ##STR160## and the indole havingthe structure: ##STR161## (vi) trimethyl amine oxide hydrate having thestructure: ##STR162## (vii) acetic acid having the structure: ##STR163##and (viii) methional having the structure: ##STR164##
 16. The enhancedfeed composition of claim 1 wherein the feed-enhancing substance is2-methyl-3-furanthiol having the structure: ##STR165##
 17. The enhancedfeed composition of claim 1 wherein the feed-enhancing substance isD-Glucosamine, an epimeric mixture of compounds having the structures:##STR166##